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    Name:
    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs
    Description:
    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs is a sterile phenol red free liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions system This low serum 2 FBS system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord Mesenchymal Stem Cell Basal Medium contains essential and non essential amino acids vitamins other organic compounds trace minerals and inorganic salts To support the proliferation and plating efficiency of various types of adult stem cells Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell specific growth kit When using this complete media system the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers extracellular matrix proteins or other substrates For mesenchymal stem cells derived from human lipoaspirates e g Adipose Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 011 or umbilical cord Umbilical Cord Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 010 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical derived MSCs Low serum ATCC PCS 500 040 For mesenchymal stem cells derived from bone marrow Bone Marrow derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 012 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Bone Marrow derived MSCs ATCC PCS 500 041 Optional media supplements Gentamicin Amphotericin B Solution ATCC PCS 999 025 Penicillin Streptomycin Amphotericin B Solution ATCC PCS 999 002 Phenol Red ATCC PCS 999 001
    Catalog Number:
    PCS-500-030
    Price:
    None
    Applications:
    Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs is a sterile, phenol red-free, liquid tissue culture medium intended for use as one component in a complete ATCC® Primary Cell Solutions™ system. This low serum (2% FBS) system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord. Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals and inorganic salts. To support the proliferation and plating efficiency of various types of adult stem cells, Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell-specific growth kit. When using this complete media system, the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers, extracellular matrix proteins or other substrates.  For mesenchymal stem cells derived from human lipoaspirates (e.g., Adipose-Derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-011) or umbilical cord (Umbilical Cord-Derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-010), supplement Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical-derived MSCs –Low serum (ATCC PCS-500-040). For mesenchymal stem cells derived from bone marrow (Bone Marrow-derived Mesenchymal Stem Cells; Normal, Human, ATCC PCS-500-012) supplement Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs with the Mesenchymal  Stem Cell Growth Kit for Bone Marrow-derived MSCs (ATCC PCS-500-041) Optional media supplements: Gentamicin-Amphotericin B Solution (ATCC PCS-999-025) Penicillin-Streptomycin-Amphotericin B Solution (ATCC PCS-999-002) Phenol Red (ATCC PCS-999-001)
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    Structured Review

    ATCC mscs
    miRNA–mRNA network in cytoprotection induced by <t>MSCs</t> . (A) MSCs-modulated network showing the interactions between miRNAs modulated by MSCs and putative targets genes interacting with at least two of these altered miRNAs, and (B) Venn diagrams depicting overlapping miRNA target genes obtained from three microRNA target prediction tools (TargetScan, miRanda, and miRDB) and a list of overlapping potential target genes for the top two differentially expressed miRNAs in the MSCs-modulated network (i.e., miR-141 and miR-377). Upregulated and downregulated miRNAs are represented, respectively, as red and green triangles. The top miRNAs in terms of fold changes had their border colors altered to yellow. Genes previously linked to fibrosis or epithelial–mesenchymal transition are represented as orange rectangles. Genes that were previously related to the TGF-β or Wnt pathways additionally had their border colors altered to purple and unrelated genes are represented in blue rectangles. Cis, cisplatin; MSCs, mesenchymal stromal cells; <t>MVs,</t> microvesicles.
    Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs is a sterile phenol red free liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions system This low serum 2 FBS system is designed to support mesenchymal stem cells derived from various normal human tissues including lipoaspirates and umbilical cord Mesenchymal Stem Cell Basal Medium contains essential and non essential amino acids vitamins other organic compounds trace minerals and inorganic salts To support the proliferation and plating efficiency of various types of adult stem cells Mesenchymal Stem Cell Basal Medium must be supplemented with the appropriate cell specific growth kit When using this complete media system the growth of undifferentiated mesenchymal stem cells is supported without the use of feeder layers extracellular matrix proteins or other substrates For mesenchymal stem cells derived from human lipoaspirates e g Adipose Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 011 or umbilical cord Umbilical Cord Derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 010 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Adipose and Umbilical derived MSCs Low serum ATCC PCS 500 040 For mesenchymal stem cells derived from bone marrow Bone Marrow derived Mesenchymal Stem Cells Normal Human ATCC PCS 500 012 supplement Mesenchymal Stem Cell Basal Medium for Adipose Umbilical and Bone Marrow derived MSCs with the Mesenchymal Stem Cell Growth Kit for Bone Marrow derived MSCs ATCC PCS 500 041 Optional media supplements Gentamicin Amphotericin B Solution ATCC PCS 999 025 Penicillin Streptomycin Amphotericin B Solution ATCC PCS 999 002 Phenol Red ATCC PCS 999 001
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    mscs - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury"

    Article Title: A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2016.00645

    miRNA–mRNA network in cytoprotection induced by MSCs . (A) MSCs-modulated network showing the interactions between miRNAs modulated by MSCs and putative targets genes interacting with at least two of these altered miRNAs, and (B) Venn diagrams depicting overlapping miRNA target genes obtained from three microRNA target prediction tools (TargetScan, miRanda, and miRDB) and a list of overlapping potential target genes for the top two differentially expressed miRNAs in the MSCs-modulated network (i.e., miR-141 and miR-377). Upregulated and downregulated miRNAs are represented, respectively, as red and green triangles. The top miRNAs in terms of fold changes had their border colors altered to yellow. Genes previously linked to fibrosis or epithelial–mesenchymal transition are represented as orange rectangles. Genes that were previously related to the TGF-β or Wnt pathways additionally had their border colors altered to purple and unrelated genes are represented in blue rectangles. Cis, cisplatin; MSCs, mesenchymal stromal cells; MVs, microvesicles.
    Figure Legend Snippet: miRNA–mRNA network in cytoprotection induced by MSCs . (A) MSCs-modulated network showing the interactions between miRNAs modulated by MSCs and putative targets genes interacting with at least two of these altered miRNAs, and (B) Venn diagrams depicting overlapping miRNA target genes obtained from three microRNA target prediction tools (TargetScan, miRanda, and miRDB) and a list of overlapping potential target genes for the top two differentially expressed miRNAs in the MSCs-modulated network (i.e., miR-141 and miR-377). Upregulated and downregulated miRNAs are represented, respectively, as red and green triangles. The top miRNAs in terms of fold changes had their border colors altered to yellow. Genes previously linked to fibrosis or epithelial–mesenchymal transition are represented as orange rectangles. Genes that were previously related to the TGF-β or Wnt pathways additionally had their border colors altered to purple and unrelated genes are represented in blue rectangles. Cis, cisplatin; MSCs, mesenchymal stromal cells; MVs, microvesicles.

    Techniques Used:

    2) Product Images from "Performance-enhanced mesenchymal stem cells via intracellular delivery of steroids"

    Article Title: Performance-enhanced mesenchymal stem cells via intracellular delivery of steroids

    Journal: Scientific Reports

    doi: 10.1038/srep04645

    Viability, metabolic activity, and morphology of budesonide treated MSC (A) MSC were plated in 96 well plates (15,000 cells/well) treated with DMSO or 0.001–100 uM budesonide and their metabolic activity was assessed by XTT at 24, 48, and 72 hours. ½ cell condition (7,500 cells/well) used as control to show a reduction in XTT signal when fewer cells are present. (B) MSCs were plated in 24 well plates (30,000 cells/well) treated with DMSO or 0.001–100 uM budesonide. At 24, 48, and 72 hours after plating, cells were harvested, and viability was measured by staining with Annexin V and propidium iodide and analyzing by flow cytometry. Staurosporine (STS) treatment for 1 and 3 hour was used as a positive control for cell death. (C) Representative phase contrast images of MSCs treated with budesonide for 48 hours show no change in morphology, density, or adherence. (D) TUNEL staining for double stranded DNA breaks after 48 hour budesonide exposure shows no increase in apoptosis. Stain control cells were treated with DNase to induce DNA breaks and served as a positive control for TUNEL stain. (DAPI, Blue; TUNEL, Green). (Bars are mean ± SEM, 2-way ANOVA with Fishers LSD test, n = 3, *p
    Figure Legend Snippet: Viability, metabolic activity, and morphology of budesonide treated MSC (A) MSC were plated in 96 well plates (15,000 cells/well) treated with DMSO or 0.001–100 uM budesonide and their metabolic activity was assessed by XTT at 24, 48, and 72 hours. ½ cell condition (7,500 cells/well) used as control to show a reduction in XTT signal when fewer cells are present. (B) MSCs were plated in 24 well plates (30,000 cells/well) treated with DMSO or 0.001–100 uM budesonide. At 24, 48, and 72 hours after plating, cells were harvested, and viability was measured by staining with Annexin V and propidium iodide and analyzing by flow cytometry. Staurosporine (STS) treatment for 1 and 3 hour was used as a positive control for cell death. (C) Representative phase contrast images of MSCs treated with budesonide for 48 hours show no change in morphology, density, or adherence. (D) TUNEL staining for double stranded DNA breaks after 48 hour budesonide exposure shows no increase in apoptosis. Stain control cells were treated with DNase to induce DNA breaks and served as a positive control for TUNEL stain. (DAPI, Blue; TUNEL, Green). (Bars are mean ± SEM, 2-way ANOVA with Fishers LSD test, n = 3, *p

    Techniques Used: Activity Assay, Staining, Flow Cytometry, Cytometry, Positive Control, TUNEL Assay

    3) Product Images from "Mesenchymal stem cells modified by FGF21 and GLP1 ameliorate lipid metabolism while reducing blood glucose in type 2 diabetic mice"

    Article Title: Mesenchymal stem cells modified by FGF21 and GLP1 ameliorate lipid metabolism while reducing blood glucose in type 2 diabetic mice

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-021-02205-z

    FGF21- and GLP1-modified MSCs could improve lipid metabolism in T2DM mice. a Hematoxylin and eosin staining of representative liver and adipose sections obtained from mice from the indicated groups (scale bars = 100 mm). b Statistics on the diameter of fat cells, * P
    Figure Legend Snippet: FGF21- and GLP1-modified MSCs could improve lipid metabolism in T2DM mice. a Hematoxylin and eosin staining of representative liver and adipose sections obtained from mice from the indicated groups (scale bars = 100 mm). b Statistics on the diameter of fat cells, * P

    Techniques Used: Modification, Mouse Assay, Staining

    FGF21- and GLP1-modified MSCs could improve glucolipid metabolism in vitro. a Quantitative real-time PCR detected the effect of FGF21- and/or GLP1-modified MSCs on the expression of srebp1c mRNA in HepG2 cells. Blank vector-modified MSCs were used as a negative control, and liraglutide was used as a positive control. The β-actin gene was used as the reference gene, ** P
    Figure Legend Snippet: FGF21- and GLP1-modified MSCs could improve glucolipid metabolism in vitro. a Quantitative real-time PCR detected the effect of FGF21- and/or GLP1-modified MSCs on the expression of srebp1c mRNA in HepG2 cells. Blank vector-modified MSCs were used as a negative control, and liraglutide was used as a positive control. The β-actin gene was used as the reference gene, ** P

    Techniques Used: Modification, In Vitro, Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation, Negative Control, Positive Control

    Transduction of MSCs with lentiviral vector particles. a Expression of EGFP in MSCs transduced at an MOI of 40 under fluorescence microscopy. Bars = 100 μm. b Flow cytometry analysis of phenotype characterization of MSC-FGF21+GLP1. The phenotypes of CD73, CD90, CD105, CD34, and CD45 expression by MSCs were detected by flow cytometry. The green lines indicate the fluorescence intensity of cells stained with the corresponding antibodies, and the red lines represent isotype-matched negative control cells. c Analysis of EGFP fluorescence by flow cytometry at 48 h after transduction at different MOIs. The MOI ranges from 0 to 55, at intervals of 5. d MSCs transduced with FGF21+GLP1 lentivirus could differentiate into osteoblasts and adipocytes. Osteogenesis was examined by alizarin red S staining for mineral nodule deposition. Adipogenesis was observed by the presence of lipid vesicles and confirmed by oil red O staining. The picture in the red box shows the enlarged observation of lipid droplets in the cell. Bars = 100 μm
    Figure Legend Snippet: Transduction of MSCs with lentiviral vector particles. a Expression of EGFP in MSCs transduced at an MOI of 40 under fluorescence microscopy. Bars = 100 μm. b Flow cytometry analysis of phenotype characterization of MSC-FGF21+GLP1. The phenotypes of CD73, CD90, CD105, CD34, and CD45 expression by MSCs were detected by flow cytometry. The green lines indicate the fluorescence intensity of cells stained with the corresponding antibodies, and the red lines represent isotype-matched negative control cells. c Analysis of EGFP fluorescence by flow cytometry at 48 h after transduction at different MOIs. The MOI ranges from 0 to 55, at intervals of 5. d MSCs transduced with FGF21+GLP1 lentivirus could differentiate into osteoblasts and adipocytes. Osteogenesis was examined by alizarin red S staining for mineral nodule deposition. Adipogenesis was observed by the presence of lipid vesicles and confirmed by oil red O staining. The picture in the red box shows the enlarged observation of lipid droplets in the cell. Bars = 100 μm

    Techniques Used: Transduction, Plasmid Preparation, Expressing, Fluorescence, Microscopy, Flow Cytometry, Staining, Negative Control

    FGF21- and GLP1-modified MSCs reduced blood glucose and weight in T2DM mice. a Four-week time course of body weight of BKS mice injected i.p. with saline (Con), MSCs, liraglutide, and FGF21- and/or GLP1-transduced MSCs. The arrow position represents the time of cell injection, n =6, * P
    Figure Legend Snippet: FGF21- and GLP1-modified MSCs reduced blood glucose and weight in T2DM mice. a Four-week time course of body weight of BKS mice injected i.p. with saline (Con), MSCs, liraglutide, and FGF21- and/or GLP1-transduced MSCs. The arrow position represents the time of cell injection, n =6, * P

    Techniques Used: Modification, Mouse Assay, Injection

    The expression of FGF21 and GLP1 in gene-modified MSCs. a Quantitative real-time PCR detected the expression of FGF21 and GLP1 mRNA in FGF21- or/and GLP1-modified cells. Unmodified MSCs and blank vector-modified MSCs were controls. The intracellular β-actin gene was used as a reference gene, ** P
    Figure Legend Snippet: The expression of FGF21 and GLP1 in gene-modified MSCs. a Quantitative real-time PCR detected the expression of FGF21 and GLP1 mRNA in FGF21- or/and GLP1-modified cells. Unmodified MSCs and blank vector-modified MSCs were controls. The intracellular β-actin gene was used as a reference gene, ** P

    Techniques Used: Expressing, Modification, Real-time Polymerase Chain Reaction, Plasmid Preparation

    4) Product Images from "Early gestational mesenchymal stem cell secretome attenuates experimental bronchopulmonary dysplasia in part via exosome-associated factor TSG-6"

    Article Title: Early gestational mesenchymal stem cell secretome attenuates experimental bronchopulmonary dysplasia in part via exosome-associated factor TSG-6

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-0903-4

    Schematic representation of MSC-CM/EXO/TSG-6 treatment regimen and outcomes in the mouse model of BPD. BPD pathology is reversed in the pups injected with secretome of hUC-derived MSCs. This therapeutic effect is mediated, at least in part, by exosomal factor TSG-6. BPD bronchopulmonary dysplasia, CM conditioned medium, EXO exosomes, hUC human umbilical cord, i.p. intraperitoneal, MSC mesenchymal stem cell, PN postnatal, TSG-6 tumor necrosis factor alpha-stimulated gene-6
    Figure Legend Snippet: Schematic representation of MSC-CM/EXO/TSG-6 treatment regimen and outcomes in the mouse model of BPD. BPD pathology is reversed in the pups injected with secretome of hUC-derived MSCs. This therapeutic effect is mediated, at least in part, by exosomal factor TSG-6. BPD bronchopulmonary dysplasia, CM conditioned medium, EXO exosomes, hUC human umbilical cord, i.p. intraperitoneal, MSC mesenchymal stem cell, PN postnatal, TSG-6 tumor necrosis factor alpha-stimulated gene-6

    Techniques Used: Injection, Derivative Assay

    Administration of EXO isolated from TSG-6 siRNA knockdown MSCs did not suppress hyperoxia-induced inflammation and associated BPD pathology. a-c Histogram showing BALF total cell count ( a ), BALF absolute neutrophil count ( b ), total BALF protein ( c ), in RA, BPD and PBS-injected, MSC-CM EXO 25 wks-injected, scr siRNA EXO 25 wks and TSG-6 siRNA EXO 25 wks-injected BPD mice at PN14. d Representative images of lung histology (H E staining) from the six experimental groups, RA (I), BPD (II), BPD + PBS (III), BPD + MSC-CM EXO 25 wks (IV), BPD + scr siRNA EXO 25 wks (V), BPD + TSG-6 siRNA EXO 25 wks (VI). Asterisks depict the increased alveolar simplification in the BPD, PBS and TSG-6 siRNA EXO 25 wks-injected BPD mice as compared to RA. ×200 magnification, Scale bar: 50 μm. e-f Histograms showing the mean chord length ( e ), alveolar area ( f ) in lungs of RA, BPD, PBS-injected, scr siRNA EXO 25 wks-injected and TSG-6 siRNA EXO 25 wks-injected BPD mice. g-h Histogram showing the RV to LV ratio ( g ) and Fulton’s index [RV/(LV + IVS)] ( h ), reflecting right ventricular hypertrophy at PN14. RA, BPD, BPD + PBS and BPD + MSC-CM EXO 25 wks samples used for comparison in this analysis are from Figs. 1 , 2 and 3 . All values are expressed as mean ± SEM; six experiments, N = 3– 8 mice per group; one-way ANOVA with Tukey’s post hoc correction; * p
    Figure Legend Snippet: Administration of EXO isolated from TSG-6 siRNA knockdown MSCs did not suppress hyperoxia-induced inflammation and associated BPD pathology. a-c Histogram showing BALF total cell count ( a ), BALF absolute neutrophil count ( b ), total BALF protein ( c ), in RA, BPD and PBS-injected, MSC-CM EXO 25 wks-injected, scr siRNA EXO 25 wks and TSG-6 siRNA EXO 25 wks-injected BPD mice at PN14. d Representative images of lung histology (H E staining) from the six experimental groups, RA (I), BPD (II), BPD + PBS (III), BPD + MSC-CM EXO 25 wks (IV), BPD + scr siRNA EXO 25 wks (V), BPD + TSG-6 siRNA EXO 25 wks (VI). Asterisks depict the increased alveolar simplification in the BPD, PBS and TSG-6 siRNA EXO 25 wks-injected BPD mice as compared to RA. ×200 magnification, Scale bar: 50 μm. e-f Histograms showing the mean chord length ( e ), alveolar area ( f ) in lungs of RA, BPD, PBS-injected, scr siRNA EXO 25 wks-injected and TSG-6 siRNA EXO 25 wks-injected BPD mice. g-h Histogram showing the RV to LV ratio ( g ) and Fulton’s index [RV/(LV + IVS)] ( h ), reflecting right ventricular hypertrophy at PN14. RA, BPD, BPD + PBS and BPD + MSC-CM EXO 25 wks samples used for comparison in this analysis are from Figs. 1 , 2 and 3 . All values are expressed as mean ± SEM; six experiments, N = 3– 8 mice per group; one-way ANOVA with Tukey’s post hoc correction; * p

    Techniques Used: Isolation, Cell Counting, Injection, Mouse Assay, Staining

    5) Product Images from "Effect of TAK1 on osteogenic differentiation of mesenchymal stem cells by regulating BMP-2 via Wnt/β-catenin and MAPK pathway"

    Article Title: Effect of TAK1 on osteogenic differentiation of mesenchymal stem cells by regulating BMP-2 via Wnt/β-catenin and MAPK pathway

    Journal: Organogenesis

    doi: 10.1080/15476278.2018.1455010

    Effect of TAK1 on the expression levels of the osteoblastic marker genes during osteogenic differentiation of MSCs. qRT-PCR analysis of the osteoblastic marker genes bone sialoprotein (BSP), osteocalcin (OSC), Alkaline phosphatase (ALP) and runt-related transcription factor 2 (RUNX2). GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p
    Figure Legend Snippet: Effect of TAK1 on the expression levels of the osteoblastic marker genes during osteogenic differentiation of MSCs. qRT-PCR analysis of the osteoblastic marker genes bone sialoprotein (BSP), osteocalcin (OSC), Alkaline phosphatase (ALP) and runt-related transcription factor 2 (RUNX2). GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p

    Techniques Used: Expressing, Marker, Quantitative RT-PCR, ALP Assay, Transfection

    p38 and JNK down regulated the effect of TAK1 on the osteogenic differentiation of MSCs. (A) The mRNA level of BMP-2 with p38 inhibitor (SB203580). (B) The mRNA level of BMP-2 with JNK inhibitor (SP600125). (C) The mRNA level of BMP-2 with p38 inhibitor (SB203580) and JNK inhibitor (SP600125). Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p
    Figure Legend Snippet: p38 and JNK down regulated the effect of TAK1 on the osteogenic differentiation of MSCs. (A) The mRNA level of BMP-2 with p38 inhibitor (SB203580). (B) The mRNA level of BMP-2 with JNK inhibitor (SP600125). (C) The mRNA level of BMP-2 with p38 inhibitor (SB203580) and JNK inhibitor (SP600125). Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p

    Techniques Used: Transfection

    Effect of TAK1 on the expression levels of β-catenin and GSK-3b during osteogenic differentiation of MSCs. qRT-PCR (A, B) and western blot (C, D) analysis of the genes and corresponding proteins. GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, or added with 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p
    Figure Legend Snippet: Effect of TAK1 on the expression levels of β-catenin and GSK-3b during osteogenic differentiation of MSCs. qRT-PCR (A, B) and western blot (C, D) analysis of the genes and corresponding proteins. GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, or added with 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Transfection

    Effect of TAK1 on the mineralization of MSCs. (A) The expression of TAK1 was downregulated by si-TAK1 and analyzed by western blot, with antibodies against TAK1 and GAPDH, respectively. (B) Mineralization was quantitated through the elution of Alizarin Red S from stained mineral deposits. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 10 and 20 days. Values are expressed as means ± S.E.M. of three independent experiments. **p
    Figure Legend Snippet: Effect of TAK1 on the mineralization of MSCs. (A) The expression of TAK1 was downregulated by si-TAK1 and analyzed by western blot, with antibodies against TAK1 and GAPDH, respectively. (B) Mineralization was quantitated through the elution of Alizarin Red S from stained mineral deposits. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 10 and 20 days. Values are expressed as means ± S.E.M. of three independent experiments. **p

    Techniques Used: Expressing, Western Blot, Staining, Transfection

    Effects of TAK1 on the expression levels of p38 and JNK during osteogenic differentiation of MSCs. (A) Western blot analysis of the protein expression levels. (B) The bar graph showed the intensities of protein expressions compared with GAPDH. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p
    Figure Legend Snippet: Effects of TAK1 on the expression levels of p38 and JNK during osteogenic differentiation of MSCs. (A) Western blot analysis of the protein expression levels. (B) The bar graph showed the intensities of protein expressions compared with GAPDH. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. * p

    Techniques Used: Expressing, Western Blot, Transfection

    Effect of TAK1 on the expression levels of TGF-β and BMP-2 during osteogenic differentiation of MSCs. qRT-PCR (A,B) and western blot (C) analysis of the genes. GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. ** p
    Figure Legend Snippet: Effect of TAK1 on the expression levels of TGF-β and BMP-2 during osteogenic differentiation of MSCs. qRT-PCR (A,B) and western blot (C) analysis of the genes. GAPDH was used as an internal control. Cells were plated at a density of 2 × 10 4 cells/cm 2 in 6-well plates. After 2 days, the medium was replaced with an osteogenic -inducing medium. Then, the cells were transfected with si-TAK1, 1 ng/ml rhTAK1 (rh1), 5 ng/ml rhTAK1 (rh5) or 50 ng/ml rhTAK1 (rh50), respectively, for 14 days. Values are expressed as means ± S.E.M. of three independent experiments. ** p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Transfection

    6) Product Images from "Mature adipocytes in bone marrow protect myeloma cells against chemotherapy through autophagy activation"

    Article Title: Mature adipocytes in bone marrow protect myeloma cells against chemotherapy through autophagy activation

    Journal: Oncotarget

    doi:

    Adipocytes produce adipokines that activate autophagy (A) Representative images of arrays show the profile of adipokine expression in the supernatants of cultured adipocytes. Medium and the supernatants from cultured MSCs or fibroblasts served as controls. We performed quantitative real-time PCR to determine the adipocyte-expressed (B) and ELISA to determine the adipocyte-secreted adipsin (C) and leptin (D) , and the expression and secretion in MSCs served as controls. (E) Western blot analysis shows the increased expression of the autophagy proteins Atg3 and LC3-I/II and unchanged β-actin in U266 cells in 24-hour culture in medium with adipocyte-conditioned media (AD-CM) or the adipokines adipsin (100 ng/ml) or leptin (25 ng/ml). Neutralizing antibodies against adipsin (2 μg/ml) or leptin (2 μg/ml), alone or in combination, were added to U266 cells in medium with adipocyte-CM. Addition of isotype IgG served as control. (F) Western blot analysis shows the reduced expression of the autophagy proteins Atg3 and LC3-I/II in U266 cells treated with the neutralizing antibodies. Antibody combination had a synergistic reduction in the autophagy protein expression as compared with treatment alone. (G) Annexin V-binding assay showed the percentage of apoptotic U266 cells in cultures with melphalan, and with or without adipsin (100 ng/ml) or leptin (25 ng/ml). (H) Annexin V-binding assay shows the percentage of apoptotic U266 cells in cultures with adipocyte-CM, with or without neutralizing antibodies in the presence or absence of melphalan. Representative results from three experiments are shown. ** P ≤ 0.01.
    Figure Legend Snippet: Adipocytes produce adipokines that activate autophagy (A) Representative images of arrays show the profile of adipokine expression in the supernatants of cultured adipocytes. Medium and the supernatants from cultured MSCs or fibroblasts served as controls. We performed quantitative real-time PCR to determine the adipocyte-expressed (B) and ELISA to determine the adipocyte-secreted adipsin (C) and leptin (D) , and the expression and secretion in MSCs served as controls. (E) Western blot analysis shows the increased expression of the autophagy proteins Atg3 and LC3-I/II and unchanged β-actin in U266 cells in 24-hour culture in medium with adipocyte-conditioned media (AD-CM) or the adipokines adipsin (100 ng/ml) or leptin (25 ng/ml). Neutralizing antibodies against adipsin (2 μg/ml) or leptin (2 μg/ml), alone or in combination, were added to U266 cells in medium with adipocyte-CM. Addition of isotype IgG served as control. (F) Western blot analysis shows the reduced expression of the autophagy proteins Atg3 and LC3-I/II in U266 cells treated with the neutralizing antibodies. Antibody combination had a synergistic reduction in the autophagy protein expression as compared with treatment alone. (G) Annexin V-binding assay showed the percentage of apoptotic U266 cells in cultures with melphalan, and with or without adipsin (100 ng/ml) or leptin (25 ng/ml). (H) Annexin V-binding assay shows the percentage of apoptotic U266 cells in cultures with adipocyte-CM, with or without neutralizing antibodies in the presence or absence of melphalan. Representative results from three experiments are shown. ** P ≤ 0.01.

    Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay

    7) Product Images from "Mesenchymal Stromal Cells Derived From Crohn's Patients Deploy Indoleamine 2,3-dioxygenase-mediated Immune Suppression, Independent of Autophagy"

    Article Title: Mesenchymal Stromal Cells Derived From Crohn's Patients Deploy Indoleamine 2,3-dioxygenase-mediated Immune Suppression, Independent of Autophagy

    Journal: Molecular Therapy

    doi: 10.1038/mt.2015.67

    Activated T cells induce autophagy on mesenchymal stromal cells (MSCs). MSCs were stimulated with indicated concentrations of ( a ) IFNγ or ( b ) TNFα for 48 hours or 4 days with ( c ) activated T cells in transwell plate. The RNA was extracted from the cells and the expression levels of Beclin, ATG12, ATG5, ATG16L1, ATG7, and LC3 mRNA was measured by the quantitative SYBR green real-time PCR. GAPDH mRNA levels were used as internal controls. Δ-Δ CT method was used to calculate the fold change. Results are plotted as mean ± SD. Similar results were obtained in a repeat experiment. MSCs and/or Hela cells were subjected to the stimulation with ( d ) Rapamycin (250 nmol/l for 16 hours) or ( e ) IFNγ (50 ng/ml for 16 hours) or ( f ) activated T cells (4 days). Cells were trypsinized and incubated with the autophagy detecting green fluorescent reagent according to the manufacturer instruction. Cells were analyzed by flow cytometry with 488 nm laser excitation and FITC detection filters. Similar results were obtained in a repeat experiment. ( g ) Cumulative mean fluorescence intensity of autophagy influx in MSCs cultured with activated T cells for 4 days in a transwell plate. MFI was plotted against unique MSC donor groups based on ATG16L1 rs2241880 A (wildtype), AG (hetero), and G (homo risk) alleles from two independent experiments done with two independent PBMC donors. Results are plotted as mean ± SD. Two-tailed T -test was performed to obtain the P values in Prism software.
    Figure Legend Snippet: Activated T cells induce autophagy on mesenchymal stromal cells (MSCs). MSCs were stimulated with indicated concentrations of ( a ) IFNγ or ( b ) TNFα for 48 hours or 4 days with ( c ) activated T cells in transwell plate. The RNA was extracted from the cells and the expression levels of Beclin, ATG12, ATG5, ATG16L1, ATG7, and LC3 mRNA was measured by the quantitative SYBR green real-time PCR. GAPDH mRNA levels were used as internal controls. Δ-Δ CT method was used to calculate the fold change. Results are plotted as mean ± SD. Similar results were obtained in a repeat experiment. MSCs and/or Hela cells were subjected to the stimulation with ( d ) Rapamycin (250 nmol/l for 16 hours) or ( e ) IFNγ (50 ng/ml for 16 hours) or ( f ) activated T cells (4 days). Cells were trypsinized and incubated with the autophagy detecting green fluorescent reagent according to the manufacturer instruction. Cells were analyzed by flow cytometry with 488 nm laser excitation and FITC detection filters. Similar results were obtained in a repeat experiment. ( g ) Cumulative mean fluorescence intensity of autophagy influx in MSCs cultured with activated T cells for 4 days in a transwell plate. MFI was plotted against unique MSC donor groups based on ATG16L1 rs2241880 A (wildtype), AG (hetero), and G (homo risk) alleles from two independent experiments done with two independent PBMC donors. Results are plotted as mean ± SD. Two-tailed T -test was performed to obtain the P values in Prism software.

    Techniques Used: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction, Incubation, Flow Cytometry, Cytometry, Fluorescence, Cell Culture, Two Tailed Test, Software

    8) Product Images from "Harnessing mesenchymal stem cell secretome: effect of extracellular matrices on pro-angiogenic signaling"

    Article Title: Harnessing mesenchymal stem cell secretome: effect of extracellular matrices on pro-angiogenic signaling

    Journal: Biotechnology and bioengineering

    doi: 10.1002/bit.27272

    Effect of ECM on pro-angiogenic signaling (A) and anti-angiogenic signaling (B) of MSCs. Comparison of the expression of these factors when MSCs (P4) were seeded on P4 dECM with (+AA) or without (−AA) ascorbic acid supplements relative to TCP (control). All factors resulting in a relative expression greater than 1.5, as indicated by the dotted line, are upregulated. Error bar Standard deviation (N=2).
    Figure Legend Snippet: Effect of ECM on pro-angiogenic signaling (A) and anti-angiogenic signaling (B) of MSCs. Comparison of the expression of these factors when MSCs (P4) were seeded on P4 dECM with (+AA) or without (−AA) ascorbic acid supplements relative to TCP (control). All factors resulting in a relative expression greater than 1.5, as indicated by the dotted line, are upregulated. Error bar Standard deviation (N=2).

    Techniques Used: Expressing, Standard Deviation

    Effect of dECM on proliferation of MSCs. MSCs were seeded on P4 dECM with (P4 ECM+AA) or without ascorbic acid (P4 ECM−AA) and cell growth was measured with respect to TCP. Error bars S.E.M (N=3). *p-value
    Figure Legend Snippet: Effect of dECM on proliferation of MSCs. MSCs were seeded on P4 dECM with (P4 ECM+AA) or without ascorbic acid (P4 ECM−AA) and cell growth was measured with respect to TCP. Error bars S.E.M (N=3). *p-value

    Techniques Used:

    Effect of dECM passage number on angiogenic signaling of MSCs. The expression of angiogenic factors when P4 MSCs were seeded on dECM generated by P3-P5 MSCs in the absence (A) and presence (B) of ascorbic acid relative to TCP (control). The concentration of pro-angiogenic (C) and anti-angiogenic (D) factors secreted by MSCs seeded on P3-P5 dECM supplemented with ascorbic acid were also normalized to the molecules secreted by MSCs seeded on dECM without ascorbic acid. The dotted lines correspond to the relative expression of 1.5. Error bar Standard deviation (N=2).
    Figure Legend Snippet: Effect of dECM passage number on angiogenic signaling of MSCs. The expression of angiogenic factors when P4 MSCs were seeded on dECM generated by P3-P5 MSCs in the absence (A) and presence (B) of ascorbic acid relative to TCP (control). The concentration of pro-angiogenic (C) and anti-angiogenic (D) factors secreted by MSCs seeded on P3-P5 dECM supplemented with ascorbic acid were also normalized to the molecules secreted by MSCs seeded on dECM without ascorbic acid. The dotted lines correspond to the relative expression of 1.5. Error bar Standard deviation (N=2).

    Techniques Used: Expressing, Generated, Concentration Assay, Standard Deviation

    Investigation of activities of factors secreted by P4 MSCs upon seeding on dECM generated by P3-P5 MSCs in the presence (+AA) and absence (−AA) of ascorbic acid. Proliferation of HUVECs in presence of conditioned medium was normalized with respect to HUVECs culture media. Error bar S.E.M (N=3). #p-value
    Figure Legend Snippet: Investigation of activities of factors secreted by P4 MSCs upon seeding on dECM generated by P3-P5 MSCs in the presence (+AA) and absence (−AA) of ascorbic acid. Proliferation of HUVECs in presence of conditioned medium was normalized with respect to HUVECs culture media. Error bar S.E.M (N=3). #p-value

    Techniques Used: Generated

    9) Product Images from "A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury"

    Article Title: A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2016.00645

    miRNA–mRNA network in cytoprotection induced by MSCs . (A) MSCs-modulated network showing the interactions between miRNAs modulated by MSCs and putative targets genes interacting with at least two of these altered miRNAs, and (B) Venn diagrams depicting overlapping miRNA target genes obtained from three microRNA target prediction tools (TargetScan, miRanda, and miRDB) and a list of overlapping potential target genes for the top two differentially expressed miRNAs in the MSCs-modulated network (i.e., miR-141 and miR-377). Upregulated and downregulated miRNAs are represented, respectively, as red and green triangles. The top miRNAs in terms of fold changes had their border colors altered to yellow. Genes previously linked to fibrosis or epithelial–mesenchymal transition are represented as orange rectangles. Genes that were previously related to the TGF-β or Wnt pathways additionally had their border colors altered to purple and unrelated genes are represented in blue rectangles. Cis, cisplatin; MSCs, mesenchymal stromal cells; MVs, microvesicles.
    Figure Legend Snippet: miRNA–mRNA network in cytoprotection induced by MSCs . (A) MSCs-modulated network showing the interactions between miRNAs modulated by MSCs and putative targets genes interacting with at least two of these altered miRNAs, and (B) Venn diagrams depicting overlapping miRNA target genes obtained from three microRNA target prediction tools (TargetScan, miRanda, and miRDB) and a list of overlapping potential target genes for the top two differentially expressed miRNAs in the MSCs-modulated network (i.e., miR-141 and miR-377). Upregulated and downregulated miRNAs are represented, respectively, as red and green triangles. The top miRNAs in terms of fold changes had their border colors altered to yellow. Genes previously linked to fibrosis or epithelial–mesenchymal transition are represented as orange rectangles. Genes that were previously related to the TGF-β or Wnt pathways additionally had their border colors altered to purple and unrelated genes are represented in blue rectangles. Cis, cisplatin; MSCs, mesenchymal stromal cells; MVs, microvesicles.

    Techniques Used:

    Related Articles

    Activity Assay:

    Article Title: Performance-enhanced mesenchymal stem cells via intracellular delivery of steroids
    Article Snippet: .. Metabolic activity, viability, and morphology assays Metabolic activity of MSCs was assessed by XTT (ATCC) following 24, 48, or 72 hour exposure to budesonide. ..

    Article Title: Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential
    Article Snippet: On day 2 and 5 after seeding, the number of EC tube formations were counted at 10× magnification by inverted microscopy and reported as n° of tube structures/field. .. Evaluation of anti-inflammatory activity of AD-MSCs and NHDFs The anti-inflammatory activity of AD-MSCs and HNDFs was tested by applying AD-MSCs-CM and HNDFs-CM on the U937 monocyte cell line (ATCC Manassas VA, USA), both in the absence and the presence of inflammatory stimuli LPS 1 μg/ml (Sigma) and TNFα 25 ng/ml (Sigma). .. The expression of the adhesion molecules CD54, CD44, CD62L and CD49d on U937 were analyzed by FC.

    Migration:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    Expressing:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    Functional Assay:

    Article Title: Intrinsic Angiogenic Potential and Migration Capacity of Human Mesenchymal Stromal Cells Derived from Menstrual Blood and Bone Marrow
    Article Snippet: .. Endothelial Tube-Like Formation Stimulated by MSC Conditioned Medium The human umbilical vein endothelial cell (HUVEC) lineage EA.hy926 (ATCC® CRL-2922™), used for endothelial tube formation and migration assay, were evaluated in regard to CD31 expression and functional properties, as shown in . .. Representative images of tubular structures formed by HUVEC incubated with mbMSC and bmMSC conditioned medium, for 48 and 72 h of culture, carried out in normoxia and 1% hypoxia, are shown in A–H.

    In Vitro:

    Article Title: A Regulatory miRNA–mRNA Network Is Associated with Tissue Repair Induced by Mesenchymal Stromal Cells in Acute Kidney Injury
    Article Snippet: A FACSCanto II flow cytometer (BD, Beckton Dickson) was used for cell acquisition, and the FlowJo software was used for data analysis. .. Coculture of MSCs or MVs with Renal Tubular Cells For in vitro assays, approximately 2 × 105 of renal epithelial tubular cells (MM55.K, ATCC® CRL-6436TM) were seeded in six-well plates (TPP, USA) and treated with nephrotoxic drug cisplatin (8 µg/mL) for 48 h, and two additional treated groups with cisplatin were co-cultured in contact with MSCs (v/v 1:1, 1 × 105 ) or MVs (50 µg/ml, sequentially each 6 h) for 48 h (Figure S4C in Supplementary Material). .. Subsequently, cells were trypsinized and subjected to analysis for apoptosis, cell proliferation, and oxidative stress analysis using the respective kits: Alexa Fluor® 488 annexin V/Dead Cell Apoptosis kit, CellTrace™ Violet Cell Proliferation kit, and MitoSOX™ Red Mitochondrial Superoxide Indicator kit (Life Technologies, USA), following the manufacturer’s recommendations (n = 6).

    Cell Culture:

    Article Title: Mesenchymal Stem/Stromal Cells in Stromal Evolution and Cancer Progression
    Article Snippet: .. Supplementary Material Cell culture: Human bone marrow mesenchymal stromal cells (MSCs) and HEK 293T cells (ATCC, Manassas, VA, USA) were cultured in α -MEM 10% FBS α -MEM powder (Invitrogen cat#12000-014, Paisley, UK), defined “FBS” (HyClone cat#SH3007003M, Logan, UT, USA) supplemented with penicillin-streptomycin (Sigma, St Louis, MO, USA), glutamine (Invitrogen cat#25030024), and minimal essential amino acids, NEAA (Invitrogen cat#11140035). .. The α -MEM powder was dissolved to commercial cell culture level water (Sigma).

    Article Title: Tumor-Activated Mesenchymal Stromal Cells Promote Osteosarcoma Stemness and Migratory Potential via IL-6 Secretion
    Article Snippet: Cell lines and sphere cultures Unless otherwise stated, all reagents were purchased from Sigma-Aldrich. .. HOS and MG63 cell lines and bone-marrow MSC were purchased from the American Type Culture Collection (ATCC) and cultured, in IMDM (Life Technologies) for tumor cells or alpha-modified Minimum Essential Medium (αMEM) for MSC plus penicillin (20U/mL), streptomycin (100 mg/mL), and 10% heat-inactivated fetal bovine serum (complete medium). ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Mesenchymal Stem/Stromal Cells in Stromal Evolution and Cancer Progression
    Article Snippet: .. Supplementary Material Cell culture: Human bone marrow mesenchymal stromal cells (MSCs) and HEK 293T cells (ATCC, Manassas, VA, USA) were cultured in α -MEM 10% FBS α -MEM powder (Invitrogen cat#12000-014, Paisley, UK), defined “FBS” (HyClone cat#SH3007003M, Logan, UT, USA) supplemented with penicillin-streptomycin (Sigma, St Louis, MO, USA), glutamine (Invitrogen cat#25030024), and minimal essential amino acids, NEAA (Invitrogen cat#11140035). .. The α -MEM powder was dissolved to commercial cell culture level water (Sigma).

    other:

    Article Title: Characterization of the antimicrobial activity of the cell-penetrating peptide TAT-RasGAP317-326
    Article Snippet: Pseudomonas aeruginosa strain PA14 was grown either in LB or BM2 medium.

    Stem Cell Culture:

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency
    Article Snippet: For Col10a1, the primer/probe cocktail was customly designed based on ref. XM_001053056.1 (Applied Biosystems, Carlsbad, CA). .. Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) . .. D1 cells were maintained with minimum Dulbecco's Modified Eagle's medium (1X DMEM, Cellgro, Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Benchmark, Gemini Bio-Products, West Sacramento, CA), and 1% penicilin-streptomicin (PS, MD Biomedicals, Thermo Fisher Scientific) under a humidified atmosphere of 5% CO2 in air, 37°C.

    Derivative Assay:

    Article Title: Circadian Rhythm and Cartilage Extracellular Matrix Genes in Osseointegration: A Genome-Wide Screening of Implant Failure by Vitamin D Deficiency
    Article Snippet: For Col10a1, the primer/probe cocktail was customly designed based on ref. XM_001053056.1 (Applied Biosystems, Carlsbad, CA). .. Bone marrow mesenchymal stem cell culture on titanium disks and vitamin D supplementation Mouse bone marrow derived mesenchymal stem cells (D1 ORL UVA [D1], ATCC® Number: CRL-s12424™) were plated at a density of 16,000 cells/well either on 48-well culture dishes (Becton Dickinson Labware, Franklin Lakes, NJ) or on sterile 10-mm Ti discs with surface of dual acid-etching and discrete crystalline deposition of HA nanoparticles (Biomet3I) . .. D1 cells were maintained with minimum Dulbecco's Modified Eagle's medium (1X DMEM, Cellgro, Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Benchmark, Gemini Bio-Products, West Sacramento, CA), and 1% penicilin-streptomicin (PS, MD Biomedicals, Thermo Fisher Scientific) under a humidified atmosphere of 5% CO2 in air, 37°C.

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    ATCC murine mesenchymal stem cell line c3h10t1 2
    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal <t>C3H10T1/2</t> cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p
    Murine Mesenchymal Stem Cell Line C3h10t1 2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC pluripotent mesenchymal precursor c2c12 cells
    Tanshinol regulates expression of KLF15 gene under condition of Dex involving glucocorticoid receptor. <t>C2C12</t> cells and MC3T3-E1 cells were treated with Dex and/or RU486 (RU, a direct target of glucocorticoid receptor) in the presence or absence of Tan for 12 h; mRNA expression of KLF15 gene was measured by qRT-PCR. Values are means ± SD of at least three independent experiments. ∗ P
    Pluripotent Mesenchymal Precursor C2c12 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC hadmsc cell culture normal hadmscs
    <t>hADMSC</t> immunophenotype and characterization of adipocyte differentiation. Positive and negative surface markers of MSCs and adipocyte differentiation marker hFATP-1 in (a). Cell cycle and G1 arrest of adipocytes in (b). hADMSC representative morphology at 4x magnification in (c) and 40x magnification in (d). Adipocyte differentiation after 14 days evidenced by Oil Red O staining. (e) and (g) are representative images at 20x and 40x magnification, respectively. Light microscopy images at day 14 of differentiation are represented in (f) and (h), corresponding to 20x and 40x magnification, respectively. The increase in the relative lipid accumulation between <t>hADMSCs</t> and adipocytes was determined by Student's t -test, ∗ p
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    ATCC mouse mesenchymal stem cell line c3h10t1 2
    GPR54 is necessary for KP-10-induced osteoblast differentiation. ( A ) Wild-type and GPR54 −/− <t>C3H10T1/2</t> cells were treated with 50 μM KP-10 for 6, 9, 12 or 24 h. Western blot analysis was performed using the indicated antibodies. BMP2 expression was normalized to β-actin expression. And the densitometry analysis was performed by ImageJ program (A, lower panel). ( B and C ) RT-PCR was performed using total RNA isolated from cultured cells. Wild-type and GPR54 −/− cells were treated with 50 μM KP-10 for 1 day ( B and C ). C.M of wild-type cells treated with 50 μM KP-10 for 12 h was collected. GPR54 −/− C3H10T1/2 cells were treated with 50 μM KP-10 or C.M for 2 days ( C ).
    Mouse Mesenchymal Stem Cell Line C3h10t1 2, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p

    Journal: bioRxiv

    Article Title: Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition

    doi: 10.1101/2021.01.19.427285

    Figure Lengend Snippet: TIE-mediated inhibition recapitulated in different in vitro and in vivo systems. (A) (To the left) The intensity of light emitted by Renilla Luciferase protein as a function of control mRNA (w/o TIE) concentration (nM). Emission of light was measured under subsaturating conditions (50-100 nM mRNA concentration). (Middle panel) The intensity of light emission by RLuc protein relative to different concentrations (50 nM and 100 nM respectively) of tested mRNA samples TIE a3, TIE a11 and control (w/o TIE). (To the right) Analysis of translation products after in vitro translation in RRL on 10% SDS PAGE to monitor RLuc expression. Visualization of protein bands was achieved by incorporation of radiolabelled 35 S-methionine, which are detected by autoradiography. (B) In vitro translation of TIE a3 and TIE a11 transcripts in the presence of m 7 G ppp G cap or a non-functional analog A ppp G. Three in vitro systems were used: Wheat Germ Extract (WGE), drosophila embryonic cell extract (S2) and HeLa cell extract. All mRNAs were translated in vitro at 50 nM concentrations and RLuc expression was normalized to control (w/o TIE) in each condition. (C) Transfection of reporter plasmids with TIE a3 or TIE a11 in two embryonic cell lines, kidney HEK293FT and mesenchymal C3H10T1/2 cell lines. Renilla luciferase expression was normalized to the control (w/o TIE). **p

    Article Snippet: Cell lines Two cell lines were used for our in vivo assays: Human embryonic kidney cell line HEK293FT (ATCC® ) and murine mesenchymal stem cell line C3H10T1/2 (clone 8, ATCC® CCL26).

    Techniques: Inhibition, In Vitro, In Vivo, Luciferase, Concentration Assay, SDS Page, Expressing, Autoradiography, Functional Assay, Transfection

    the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p

    Journal: bioRxiv

    Article Title: Translation Inhibitory Elements from Hox a3 and a11 mRNAs use uORFs for translation inhibition

    doi: 10.1101/2021.01.19.427285

    Figure Lengend Snippet: the uAUG111 in TIE a3 is translated through 5’UTR of Hox a3. (A) Three transcripts were used for this experiment: full length 5’UTR of Hox a3, a deletion mutant at nucleotide G333 in IRES a3 and a control transcript without TIE. To test the translation of uORF in TIE a3 starting form uAUG111, a deletion of G in IRES a3 at position 333 was performed to create a longer uORF that is in the same frame as the ORF of Renilla luciferase to create an N-terminally extended luciferase. Transcripts were translated in vitro in RRL and products were loaded on 10% SDS-PAGE in the presence of 35 S-Methionine (B) In vivo luciferase assays in two embryonic cells lines; HEK293FT (left) and C3H10T1/2 (right). Reporter constructs in pmirGlo containing TIE a3, u(AUG/UAC)111, and without TIE, were transfected in the two indicated cell lines. Renilla luciferase expression was normalized to the control (w/o TIE), which was set to 100%. **p

    Article Snippet: Cell lines Two cell lines were used for our in vivo assays: Human embryonic kidney cell line HEK293FT (ATCC® ) and murine mesenchymal stem cell line C3H10T1/2 (clone 8, ATCC® CCL26).

    Techniques: Mutagenesis, Luciferase, In Vitro, SDS Page, In Vivo, Construct, Transfection, Expressing

    Tanshinol regulates expression of KLF15 gene under condition of Dex involving glucocorticoid receptor. C2C12 cells and MC3T3-E1 cells were treated with Dex and/or RU486 (RU, a direct target of glucocorticoid receptor) in the presence or absence of Tan for 12 h; mRNA expression of KLF15 gene was measured by qRT-PCR. Values are means ± SD of at least three independent experiments. ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Tanshinol Rescues the Impaired Bone Formation Elicited by Glucocorticoid Involved in KLF15 Pathway

    doi: 10.1155/2016/1092746

    Figure Lengend Snippet: Tanshinol regulates expression of KLF15 gene under condition of Dex involving glucocorticoid receptor. C2C12 cells and MC3T3-E1 cells were treated with Dex and/or RU486 (RU, a direct target of glucocorticoid receptor) in the presence or absence of Tan for 12 h; mRNA expression of KLF15 gene was measured by qRT-PCR. Values are means ± SD of at least three independent experiments. ∗ P

    Article Snippet: Cell Culture and Osteoblastic Differentiation Assay The pluripotent mesenchymal precursor C2C12 cells and preosteoblastic MC3T3-E1 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Expressing, Quantitative RT-PCR

    Tanshinol attenuates downregulation of canonical Wnt signaling elicited by Dex associated with regulation KLF15. ((a) and (b)) C2C12 cells or MC3T3-E1 cells were cotransfected with the Tcf4-luc or FoxO3a-luc reporter plasmid in combination with KLF15 siRNA or the scrambled sequence. ((c) and (d)) C2C12 cells or MC3T3-E1 cells were infected with the FoxO3a-luc or Tcf4-luc reporter plasmid in combination with recombinant adenovirus Ad-KLF15 or mock (noninfection). Luciferase activity assays were explored using the Dual-Luciferase Reporter Assay System as described under Section 2.8 in Materials and Methods. The data represent mean ± SD of luciferase relative luminescence units (RLU) normalized to corresponding renilla luciferase activity (triplicates). (e) Tcf4 (a requisite mediator for downstream effector Tcf of canonical Wnt pathway contributing to bone formation) in MC3T3-E1 cells exposed to Ad-KLF15 and mock was detected by Western blot assay. Representative figure was shown on the left panel, and quantification is shown on the right panel. Bars indicate mean ± SD of triplicate determinations. ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Tanshinol Rescues the Impaired Bone Formation Elicited by Glucocorticoid Involved in KLF15 Pathway

    doi: 10.1155/2016/1092746

    Figure Lengend Snippet: Tanshinol attenuates downregulation of canonical Wnt signaling elicited by Dex associated with regulation KLF15. ((a) and (b)) C2C12 cells or MC3T3-E1 cells were cotransfected with the Tcf4-luc or FoxO3a-luc reporter plasmid in combination with KLF15 siRNA or the scrambled sequence. ((c) and (d)) C2C12 cells or MC3T3-E1 cells were infected with the FoxO3a-luc or Tcf4-luc reporter plasmid in combination with recombinant adenovirus Ad-KLF15 or mock (noninfection). Luciferase activity assays were explored using the Dual-Luciferase Reporter Assay System as described under Section 2.8 in Materials and Methods. The data represent mean ± SD of luciferase relative luminescence units (RLU) normalized to corresponding renilla luciferase activity (triplicates). (e) Tcf4 (a requisite mediator for downstream effector Tcf of canonical Wnt pathway contributing to bone formation) in MC3T3-E1 cells exposed to Ad-KLF15 and mock was detected by Western blot assay. Representative figure was shown on the left panel, and quantification is shown on the right panel. Bars indicate mean ± SD of triplicate determinations. ∗ P

    Article Snippet: Cell Culture and Osteoblastic Differentiation Assay The pluripotent mesenchymal precursor C2C12 cells and preosteoblastic MC3T3-E1 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Plasmid Preparation, Sequencing, Infection, Recombinant, Luciferase, Activity Assay, Reporter Assay, Western Blot

    Tanshinol counteracts inhibition of osteoblastic differentiation and bone formation elicited by Dex in connection with downregulation of KLF15. C2C12 cells and MC3T3-E1 cells were transfected with KLF15 siRNA for 18 h, followed by DMEM medium supplemented with Dex in the presence or absence of tanshinol for 7 days. (a) Capacity of osteoblastic differentiation in C2C12 cells was determined by using ALP staining. (b) Capacity of osteoblastic differentiation in MC3T3-E1 cells. Original magnification (×100) in representative microscopic images. (c) Effects of knockdown of KLF15 on activity of bone formation. MC3T3-E1 cells were treated with KLF15 siRNA for 18 h, followed by Dex treatment with or without tanshinol for 21 days. Mineralization activity with the indicated treatments was stained using Alizarin Red S at day 21. Original magnification (×100) in representative microscopic images (upper panel). Quantitative determination was carried out by CPC solution (pH 7.0) (lower panel). Vehicle: vehicle control (Veh). Values are means ± SD of at least three independent experiments. ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Tanshinol Rescues the Impaired Bone Formation Elicited by Glucocorticoid Involved in KLF15 Pathway

    doi: 10.1155/2016/1092746

    Figure Lengend Snippet: Tanshinol counteracts inhibition of osteoblastic differentiation and bone formation elicited by Dex in connection with downregulation of KLF15. C2C12 cells and MC3T3-E1 cells were transfected with KLF15 siRNA for 18 h, followed by DMEM medium supplemented with Dex in the presence or absence of tanshinol for 7 days. (a) Capacity of osteoblastic differentiation in C2C12 cells was determined by using ALP staining. (b) Capacity of osteoblastic differentiation in MC3T3-E1 cells. Original magnification (×100) in representative microscopic images. (c) Effects of knockdown of KLF15 on activity of bone formation. MC3T3-E1 cells were treated with KLF15 siRNA for 18 h, followed by Dex treatment with or without tanshinol for 21 days. Mineralization activity with the indicated treatments was stained using Alizarin Red S at day 21. Original magnification (×100) in representative microscopic images (upper panel). Quantitative determination was carried out by CPC solution (pH 7.0) (lower panel). Vehicle: vehicle control (Veh). Values are means ± SD of at least three independent experiments. ∗ P

    Article Snippet: Cell Culture and Osteoblastic Differentiation Assay The pluripotent mesenchymal precursor C2C12 cells and preosteoblastic MC3T3-E1 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Inhibition, Transfection, ALP Assay, Staining, Activity Assay

    hADMSC immunophenotype and characterization of adipocyte differentiation. Positive and negative surface markers of MSCs and adipocyte differentiation marker hFATP-1 in (a). Cell cycle and G1 arrest of adipocytes in (b). hADMSC representative morphology at 4x magnification in (c) and 40x magnification in (d). Adipocyte differentiation after 14 days evidenced by Oil Red O staining. (e) and (g) are representative images at 20x and 40x magnification, respectively. Light microscopy images at day 14 of differentiation are represented in (f) and (h), corresponding to 20x and 40x magnification, respectively. The increase in the relative lipid accumulation between hADMSCs and adipocytes was determined by Student's t -test, ∗ p

    Journal: Stem Cells International

    Article Title: Hydrogen Peroxide-Induced DNA Damage and Repair through the Differentiation of Human Adipose-Derived Mesenchymal Stem Cells

    doi: 10.1155/2018/1615497

    Figure Lengend Snippet: hADMSC immunophenotype and characterization of adipocyte differentiation. Positive and negative surface markers of MSCs and adipocyte differentiation marker hFATP-1 in (a). Cell cycle and G1 arrest of adipocytes in (b). hADMSC representative morphology at 4x magnification in (c) and 40x magnification in (d). Adipocyte differentiation after 14 days evidenced by Oil Red O staining. (e) and (g) are representative images at 20x and 40x magnification, respectively. Light microscopy images at day 14 of differentiation are represented in (f) and (h), corresponding to 20x and 40x magnification, respectively. The increase in the relative lipid accumulation between hADMSCs and adipocytes was determined by Student's t -test, ∗ p

    Article Snippet: 2.1. hADMSC Cell Culture Normal hADMSCs (ATCC® PCS-500-011™) were cultured at an early passage (passage 4).

    Techniques: Marker, Staining, Light Microscopy

    GPR54 is necessary for KP-10-induced osteoblast differentiation. ( A ) Wild-type and GPR54 −/− C3H10T1/2 cells were treated with 50 μM KP-10 for 6, 9, 12 or 24 h. Western blot analysis was performed using the indicated antibodies. BMP2 expression was normalized to β-actin expression. And the densitometry analysis was performed by ImageJ program (A, lower panel). ( B and C ) RT-PCR was performed using total RNA isolated from cultured cells. Wild-type and GPR54 −/− cells were treated with 50 μM KP-10 for 1 day ( B and C ). C.M of wild-type cells treated with 50 μM KP-10 for 12 h was collected. GPR54 −/− C3H10T1/2 cells were treated with 50 μM KP-10 or C.M for 2 days ( C ).

    Journal: Scientific Reports

    Article Title: Kisspeptin-10 (KP-10) stimulates osteoblast differentiation through GPR54-mediated regulation of BMP2 expression and activation

    doi: 10.1038/s41598-018-20571-2

    Figure Lengend Snippet: GPR54 is necessary for KP-10-induced osteoblast differentiation. ( A ) Wild-type and GPR54 −/− C3H10T1/2 cells were treated with 50 μM KP-10 for 6, 9, 12 or 24 h. Western blot analysis was performed using the indicated antibodies. BMP2 expression was normalized to β-actin expression. And the densitometry analysis was performed by ImageJ program (A, lower panel). ( B and C ) RT-PCR was performed using total RNA isolated from cultured cells. Wild-type and GPR54 −/− cells were treated with 50 μM KP-10 for 1 day ( B and C ). C.M of wild-type cells treated with 50 μM KP-10 for 12 h was collected. GPR54 −/− C3H10T1/2 cells were treated with 50 μM KP-10 or C.M for 2 days ( C ).

    Article Snippet: Cell culture The mouse mesenchymal stem cell line C3H10T1/2 (ATCC, Manassas, VA) was maintained in DMEM containing 10% FBS, 100 units/mL penicillin and 100 μg/mL streptomycin in humidified air containing 5% CO2 at 37 °C.

    Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Cell Culture

    KP-10 induces osteogenic gene expressions in C3H10T1/2 cells. ( A and B ) RT-PCR were performed using total RNA isolated from C3H10T1/2 cells treated with 5 or 50 μM KP-10 for 2 days ( A ), 50 μM KP-10 for 1 or 2 days ( B ). ( C ) Real-time PCR were performed using total RNA isolated cells treated with 50 μM of KP-10 for 2 days. * p

    Journal: Scientific Reports

    Article Title: Kisspeptin-10 (KP-10) stimulates osteoblast differentiation through GPR54-mediated regulation of BMP2 expression and activation

    doi: 10.1038/s41598-018-20571-2

    Figure Lengend Snippet: KP-10 induces osteogenic gene expressions in C3H10T1/2 cells. ( A and B ) RT-PCR were performed using total RNA isolated from C3H10T1/2 cells treated with 5 or 50 μM KP-10 for 2 days ( A ), 50 μM KP-10 for 1 or 2 days ( B ). ( C ) Real-time PCR were performed using total RNA isolated cells treated with 50 μM of KP-10 for 2 days. * p

    Article Snippet: Cell culture The mouse mesenchymal stem cell line C3H10T1/2 (ATCC, Manassas, VA) was maintained in DMEM containing 10% FBS, 100 units/mL penicillin and 100 μg/mL streptomycin in humidified air containing 5% CO2 at 37 °C.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Real-time Polymerase Chain Reaction