Structured Review

Nichirei fetal bovine serum fbs
Migration activity of EMT-induced cancer cells. (A and B) To measure cell migration activity, A549 (left panels) and Panc-1 (right panels) cells were subjected to the in vitro wound healing assay in the presence or absence (Control) of TGF-ß, as described in the text. Phase-contrast micrographs of the cultures were taken after 16-h incubation. Black broken lines indicate initial wound edges. Scale bar, 50 µm. B) Cell migration area was estimated by analyzing the pixel with Image J. Each bar indicates the mean ± SD for the areas of migrated cells in 3 different fields (right panels). (C) Cells that had been pretreated without or with TGF-ß for 24 h were incubated in 1% <t>FBS-containing</t> medium without or with TGF-ß onto a 24-well plate for 6 h at <t>37°C.</t> The cell migration was monitored with a time-lapse video system for 12 h. Cell migration distance was measured for 15 randomly selected cells. Each bar indicates the mean ± SD for cell migration velocity of 15 cells. Other experimental conditions are described in Materials and Methods .
Fetal Bovine Serum Fbs, supplied by Nichirei, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fetal bovine serum fbs/product/Nichirei
Average 92 stars, based on 3 article reviews
Price from $9.99 to $1999.99
fetal bovine serum fbs - by Bioz Stars, 2020-07
92/100 stars

Images

1) Product Images from "Epithelial-Mesenchymal Transition Stimulates Human Cancer Cells to Extend Microtubule-based Invasive Protrusions and Suppresses Cell Growth in Collagen Gel"

Article Title: Epithelial-Mesenchymal Transition Stimulates Human Cancer Cells to Extend Microtubule-based Invasive Protrusions and Suppresses Cell Growth in Collagen Gel

Journal: PLoS ONE

doi: 10.1371/journal.pone.0053209

Migration activity of EMT-induced cancer cells. (A and B) To measure cell migration activity, A549 (left panels) and Panc-1 (right panels) cells were subjected to the in vitro wound healing assay in the presence or absence (Control) of TGF-ß, as described in the text. Phase-contrast micrographs of the cultures were taken after 16-h incubation. Black broken lines indicate initial wound edges. Scale bar, 50 µm. B) Cell migration area was estimated by analyzing the pixel with Image J. Each bar indicates the mean ± SD for the areas of migrated cells in 3 different fields (right panels). (C) Cells that had been pretreated without or with TGF-ß for 24 h were incubated in 1% FBS-containing medium without or with TGF-ß onto a 24-well plate for 6 h at 37°C. The cell migration was monitored with a time-lapse video system for 12 h. Cell migration distance was measured for 15 randomly selected cells. Each bar indicates the mean ± SD for cell migration velocity of 15 cells. Other experimental conditions are described in Materials and Methods .
Figure Legend Snippet: Migration activity of EMT-induced cancer cells. (A and B) To measure cell migration activity, A549 (left panels) and Panc-1 (right panels) cells were subjected to the in vitro wound healing assay in the presence or absence (Control) of TGF-ß, as described in the text. Phase-contrast micrographs of the cultures were taken after 16-h incubation. Black broken lines indicate initial wound edges. Scale bar, 50 µm. B) Cell migration area was estimated by analyzing the pixel with Image J. Each bar indicates the mean ± SD for the areas of migrated cells in 3 different fields (right panels). (C) Cells that had been pretreated without or with TGF-ß for 24 h were incubated in 1% FBS-containing medium without or with TGF-ß onto a 24-well plate for 6 h at 37°C. The cell migration was monitored with a time-lapse video system for 12 h. Cell migration distance was measured for 15 randomly selected cells. Each bar indicates the mean ± SD for cell migration velocity of 15 cells. Other experimental conditions are described in Materials and Methods .

Techniques Used: Migration, Activity Assay, In Vitro, Wound Healing Assay, Incubation

2) Product Images from "Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern"

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern

Journal: The Journal of Veterinary Medical Science

doi: 10.1292/jvms.18-0202

Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.
Figure Legend Snippet: Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.

Techniques Used: Cell Culture, Derivative Assay

3) Product Images from "Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern"

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern

Journal: The Journal of Veterinary Medical Science

doi: 10.1292/jvms.18-0202

Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.
Figure Legend Snippet: Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.

Techniques Used: Cell Culture, Derivative Assay

4) Product Images from "Functional analysis of fatty acid binding protein 7 and its effect on fatty acid of renal cell carcinoma cell lines"

Article Title: Functional analysis of fatty acid binding protein 7 and its effect on fatty acid of renal cell carcinoma cell lines

Journal: BMC Cancer

doi: 10.1186/s12885-017-3184-x

Effect of FABP7 on cell proliferation and cell cycle of TUHR14TKB cells. TUHR14TKB cells were cultured in RPMI 1640 medium containing 10% FBS, 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. Assays to determine the rate of cell proliferation: a MTS assay. The data represent the average and standard deviation (error bars) of five experiments. b Cell counts. The data represent the average and standard deviation (error bars) of four experiments. c The stages of the cell cycles of TUHR14TKB FABP7 and TUBR14TKB lacZ were determined using flow cytometry
Figure Legend Snippet: Effect of FABP7 on cell proliferation and cell cycle of TUHR14TKB cells. TUHR14TKB cells were cultured in RPMI 1640 medium containing 10% FBS, 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. Assays to determine the rate of cell proliferation: a MTS assay. The data represent the average and standard deviation (error bars) of five experiments. b Cell counts. The data represent the average and standard deviation (error bars) of four experiments. c The stages of the cell cycles of TUHR14TKB FABP7 and TUBR14TKB lacZ were determined using flow cytometry

Techniques Used: Cell Culture, MTS Assay, Standard Deviation, Flow Cytometry, Cytometry

Effect of FABP7 on the migration of TUHR14TKB cells. TUHR14TKB cells were transfected with the FABP7 or lacZ expression vector. a Western blot analysis of FABP7 expression by cells transfected with the FABP7 vector or control (lacZ) vector. b Real-time PCR analysis of FABP7 expression in cells transfected with the FABP7 vector or lacZ vector. c Wound-healing assays. TUHR14TKB transfectants were cultured in RPMI 1640 medium containing with 10% FBS or 1% FBS with 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. The data represent the average and standard deviation (error bars) of four experiments
Figure Legend Snippet: Effect of FABP7 on the migration of TUHR14TKB cells. TUHR14TKB cells were transfected with the FABP7 or lacZ expression vector. a Western blot analysis of FABP7 expression by cells transfected with the FABP7 vector or control (lacZ) vector. b Real-time PCR analysis of FABP7 expression in cells transfected with the FABP7 vector or lacZ vector. c Wound-healing assays. TUHR14TKB transfectants were cultured in RPMI 1640 medium containing with 10% FBS or 1% FBS with 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. The data represent the average and standard deviation (error bars) of four experiments

Techniques Used: Migration, Transfection, Expressing, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Standard Deviation

Expression of FABP7 during subculture of TUHR14TKB cells. The zero passage (0) was started when the cells were received from RIKEN. TUHR14TKB cells were cultured in RPMI 1640 medium containing 10% FBS for one to two weeks, harvested when they reached confluence, and assayed for FABP7 expression. a Western blot analysis of FABP7 expression. b Real-time PCR analysis of FABP7 expression
Figure Legend Snippet: Expression of FABP7 during subculture of TUHR14TKB cells. The zero passage (0) was started when the cells were received from RIKEN. TUHR14TKB cells were cultured in RPMI 1640 medium containing 10% FBS for one to two weeks, harvested when they reached confluence, and assayed for FABP7 expression. a Western blot analysis of FABP7 expression. b Real-time PCR analysis of FABP7 expression

Techniques Used: Expressing, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction

Effect of fatty acids on cell proliferation. a Docosatetraenoic acid or EPA concentration of TUHR14TKB cells transfected with the FABP7 or lacZ expression vector. Four independent cell cultures were harvested, and the concentration of docosatetraenoic acid or EPA was determined. The data represent the average and standard deviation. b Cell proliferation was assayed in the presence or absence of 50 μM docosatetraenoic acid or EPA in RPMI 1640 medium containing 10% FBS, 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. The doubling times of TUHR14TKB lacZ cells were determined using an MTS assay. The data represent the average and standard deviation (error bars) of 12 experiments
Figure Legend Snippet: Effect of fatty acids on cell proliferation. a Docosatetraenoic acid or EPA concentration of TUHR14TKB cells transfected with the FABP7 or lacZ expression vector. Four independent cell cultures were harvested, and the concentration of docosatetraenoic acid or EPA was determined. The data represent the average and standard deviation. b Cell proliferation was assayed in the presence or absence of 50 μM docosatetraenoic acid or EPA in RPMI 1640 medium containing 10% FBS, 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate. The doubling times of TUHR14TKB lacZ cells were determined using an MTS assay. The data represent the average and standard deviation (error bars) of 12 experiments

Techniques Used: Concentration Assay, Transfection, Expressing, Plasmid Preparation, Standard Deviation, MTS Assay

Related Articles

Modification:

Article Title: Induced overexpression of CD44 associated with resistance to apoptosis on DNA damage response in human head and neck squamous cell carcinoma cells
Article Snippet: .. Dulbecco's modified Eagle's medium (DMEM) was purchased from Invitrogen (Carlsbad, CA, USA), and fetal bovine serum (FBS) was purchased from Nichirei Bioscience (Tokyo, Japan). .. Primary antibodies against CD44 and cPARP were purchased from Cell Signaling Technology (Danvers, MA, USA), and primary antibodies against phospho-CHK1 (S301) and β-actin were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern
Article Snippet: .. After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/m l of penicillin and 0.1 mg/m l of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days. ..

Incubation:

Article Title: A Novel Method to Apply Osteogenic Potential of Adipose Derived Stem Cells in Orthopaedic Surgery
Article Snippet: .. Preparing hADSCs The hADSCs were suspended in DMEM (Wako Pure Chemical Industries, Ltd.) supplemented with 10% fetal bovine serum (FBS) (NICHIREI BIOSCIENCES, INC.) and 1% Penicillin-Streptomycin Solution (P/S) (Wako Pure Chemical Industries, Ltd.), and incubated at 37°C for 1day. ..

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern
Article Snippet: .. After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/ml of penicillin and 0.1 mg/ml of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days. ..

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern
Article Snippet: .. After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/m l of penicillin and 0.1 mg/m l of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days. ..

Recombinant:

Article Title: Srf destabilizes cellular identity by suppressing cell-type-specific gene expression programs
Article Snippet: .. These cells were maintained on a type IV collagen-coated culture plate in a medium consisting of a 1:1 mixture of DMEM/Ham’s F-12 with l -glutamine, HEPES, and sodium pyruvate (Wako) supplemented with 10% fetal bovine serum (FBS) (vol/vol) (Nichirei Bioscience), 1 µg/ml insulin (Wako), 100 nM dexamethasone (Sigma-Aldrich), 10 mM nicotinamide (Sigma-Aldrich), 50 µM β-mercaptoethanol (Nacalai Tesque), 50 ng/ml human recombinant hepatocyte growth factor (HGF; Peprotech), and 20 ng/ml EGF. .. Mouse UBCs (from Kenji Osafune) were maintained on a gelatin-coated culture plate in DMEM (Wako) supplemented with 10% FBS (Thermo Fisher Scientific) and 1 mM sodium pyruvate (Nacalai Tesque).

Centrifugation:

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern
Article Snippet: .. After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/ml of penicillin and 0.1 mg/ml of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days. ..

Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern
Article Snippet: .. After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/m l of penicillin and 0.1 mg/m l of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days. ..

Cell Culture:

Article Title: Epithelial-Mesenchymal Transition Stimulates Human Cancer Cells to Extend Microtubule-based Invasive Protrusions and Suppresses Cell Growth in Collagen Gel
Article Snippet: .. These cell lines were cultured in DMEM/F12 medium (Invitrogen, Carlsbed, CA) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences, Tokyo) at 37°C in a humidified atmosphere of 5% CO2 and 95% air. ..

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    Nichirei fbs
    Islet cell sheet morphology after fiber-modified adenoviral vector infection. Following infection with AdK7-CA-GFP at the indicated MOIs, the islet cells were incubated in <t>RPMI</t> 1640 with 10% <t>FBS</t> for 96 h and subjected to phase-contrast microscopic analysis. Scale bars: 50 μm.
    Fbs, supplied by Nichirei, used in various techniques. Bioz Stars score: 92/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fbs/product/Nichirei
    Average 92 stars, based on 74 article reviews
    Price from $9.99 to $1999.99
    fbs - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    92
    Nichirei msc like cell induction medium
    Recapitulation of FOP phenotype shown by two chondrogenesis pathways. (A) Schematic view of a protocol for <t>MSC-like</t> cell induction. Induced SM were treated with FGF2 (4 ng/ml) and <t>FBS</t> (10%) for 12 days. (B) FACS analysis using surface markers for MSCs at day 12 of MSC-like cell induction. CD44 + , CD73 + , CD105 + and CD45 (PTPRC) − cells were induced. SM cells were used as control populations. (C) Schematic view of MSC-like cell-CI and SCL-CI using FOP-iPSCs and resFOP-iPSCs. Briefly, induced MSC-like cells and SCL were spotted onto fibronectin-coated dishes and treated with CI basal media supplemented with activin A (30 ng/ml) for 5 days. (D) ACVR1 expression in MSC-like cells and SCL induced from both FOP-iPSCs and resFOP-iPSCs was assessed by RT-qPCR. Error bars represent s.e.m. ( n =6). The expression level of resFOP-MSCs was set to 1. (E-G) Evaluation of MSC-like cell-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (E), Alcian Blue staining (F) and GAG/DNA analysis (G). (H-J) Evaluation of SCL-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (H), Alcian Blue staining (I) and GAG/DNA analysis (J). (K,L) Evaluation of R667 and Rapamycin efficacy on MSC-like cell-CI. Chondrogenic differentiation was assessed at day 5 of CI by Alcian Blue staining (K) and GAG/DNA analysis (L). (M-P) In vitro study regarding cell-of-origins of the ectopic bone observed in FOP. PDGFRα + /CD31 – and PDGFRα – /CD31 – populations were isolated by FACS (M). Chondrogenic potential of each population was assessed at day 5 of CI by GAG/DNA (N) and RT-qPCR analysis (O). Expression levels of PAI1 and MMP1 , both surrogate markers of aberrant FOP-ACVR1 signaling, were higher in PDGFRα + /CD31 – cells (P). Error bars represent s.e.m. ( n =3). * P
    Msc Like Cell Induction Medium, supplied by Nichirei, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/msc like cell induction medium/product/Nichirei
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    msc like cell induction medium - by Bioz Stars, 2020-07
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    Islet cell sheet morphology after fiber-modified adenoviral vector infection. Following infection with AdK7-CA-GFP at the indicated MOIs, the islet cells were incubated in RPMI 1640 with 10% FBS for 96 h and subjected to phase-contrast microscopic analysis. Scale bars: 50 μm.

    Journal: Cell Medicine

    Article Title: Efficient Gene Transduction of Dispersed Islet Cells in Culture Using Fiber-Modified Adenoviral Vectors

    doi: 10.3727/215517915X689083

    Figure Lengend Snippet: Islet cell sheet morphology after fiber-modified adenoviral vector infection. Following infection with AdK7-CA-GFP at the indicated MOIs, the islet cells were incubated in RPMI 1640 with 10% FBS for 96 h and subjected to phase-contrast microscopic analysis. Scale bars: 50 μm.

    Article Snippet: Isolated islets were cultured overnight in Roswell Park Memorial Institute (RPMI)-1640 medium (Sigma-Aldrich) containing 10% FBS (Nichirei Biosciences, Tokyo, Japan) and 5.5 mM glucose at 37°C and were subsequently treated with trypsin–ethylenediaminetetraacetic acid (EDTA) (Invitrogen, Carlsbad, CA, USA) for dispersion into single islet cells ( ).

    Techniques: Modification, Plasmid Preparation, Infection, Incubation

    Scheme of the experimental procedure. Pancreatic islets were isolated from male Lewis rats 10- to 12-weeks old and were cultured overnight in Roswell Park Memorial Institute (RPMI)-1640 medium containing 10% fetal bovine serum (FBS) and 5.5 mM glucose at 37°C. Dispersed islet cells were harvested by treatment of the cultured islets with trypsin-ethylenediaminetetraacetic acid (EDTA) and were plated onto laminin-5-coated temperature-responsive culture dishes (24 h). At 0 h, a fully confluent monolayer of islet cells was obtained and was infected with adenoviral (Ad) vectors. Briefly, the medium was replenished with RPMI-1640 medium containing 2% FBS, and then the cells were exposed to normal Ad-CA-green fluorescent protein (GFP) or fiber-modified (AdK7-CA-GFP) vectors at multiplicities of infection (MOIs) of 0–30 in 100 µl of the medium for 1 h at 37°C. Following two washes with RPMI-1640 with 10% FBS, the islet cells were incubated in the medium for either 48 h for fluorescence-activated cell sorting (FACS) analysis or 96 h for the assessment of cytotoxicity [lactate dehydrogenase (LDH) assay]. The medium for culturing for 96 h was renewed at 48 h following washing of the cells twice with the medium.

    Journal: Cell Medicine

    Article Title: Efficient Gene Transduction of Dispersed Islet Cells in Culture Using Fiber-Modified Adenoviral Vectors

    doi: 10.3727/215517915X689083

    Figure Lengend Snippet: Scheme of the experimental procedure. Pancreatic islets were isolated from male Lewis rats 10- to 12-weeks old and were cultured overnight in Roswell Park Memorial Institute (RPMI)-1640 medium containing 10% fetal bovine serum (FBS) and 5.5 mM glucose at 37°C. Dispersed islet cells were harvested by treatment of the cultured islets with trypsin-ethylenediaminetetraacetic acid (EDTA) and were plated onto laminin-5-coated temperature-responsive culture dishes (24 h). At 0 h, a fully confluent monolayer of islet cells was obtained and was infected with adenoviral (Ad) vectors. Briefly, the medium was replenished with RPMI-1640 medium containing 2% FBS, and then the cells were exposed to normal Ad-CA-green fluorescent protein (GFP) or fiber-modified (AdK7-CA-GFP) vectors at multiplicities of infection (MOIs) of 0–30 in 100 µl of the medium for 1 h at 37°C. Following two washes with RPMI-1640 with 10% FBS, the islet cells were incubated in the medium for either 48 h for fluorescence-activated cell sorting (FACS) analysis or 96 h for the assessment of cytotoxicity [lactate dehydrogenase (LDH) assay]. The medium for culturing for 96 h was renewed at 48 h following washing of the cells twice with the medium.

    Article Snippet: Isolated islets were cultured overnight in Roswell Park Memorial Institute (RPMI)-1640 medium (Sigma-Aldrich) containing 10% FBS (Nichirei Biosciences, Tokyo, Japan) and 5.5 mM glucose at 37°C and were subsequently treated with trypsin–ethylenediaminetetraacetic acid (EDTA) (Invitrogen, Carlsbad, CA, USA) for dispersion into single islet cells ( ).

    Techniques: Isolation, Cell Culture, Infection, Modification, Incubation, Fluorescence, FACS, Lactate Dehydrogenase Assay

    Transduction efficiency of GFP gene into dispersed islet cells by adenoviral vectors. (A) Representative histograms of GFP expression in dispersed islet cells infected with adenoviral vectors. Dispersed islet cells in 24-well plates were infected with Ad-CA-GFP (top) or AdK7-CA-GFP (bottom) at the indicated MOIs. The cells were further cultured in RPMI-1640 with 10% FBS for 47 h and were analyzed by flow cytometry. (B, C) The percentage (B) and mean fluorescence intensity (MFI) (C) of GFP-positive islet cells were determined by flow cytometry. Data are presented as means ± standard deviation ( n = 3). The asterisks indicate significant differences between Ad-CA-GFP (gray bars) and AdK7-CA-GFP (black bars) determined by Sidak’s multiple comparisons test ( p

    Journal: Cell Medicine

    Article Title: Efficient Gene Transduction of Dispersed Islet Cells in Culture Using Fiber-Modified Adenoviral Vectors

    doi: 10.3727/215517915X689083

    Figure Lengend Snippet: Transduction efficiency of GFP gene into dispersed islet cells by adenoviral vectors. (A) Representative histograms of GFP expression in dispersed islet cells infected with adenoviral vectors. Dispersed islet cells in 24-well plates were infected with Ad-CA-GFP (top) or AdK7-CA-GFP (bottom) at the indicated MOIs. The cells were further cultured in RPMI-1640 with 10% FBS for 47 h and were analyzed by flow cytometry. (B, C) The percentage (B) and mean fluorescence intensity (MFI) (C) of GFP-positive islet cells were determined by flow cytometry. Data are presented as means ± standard deviation ( n = 3). The asterisks indicate significant differences between Ad-CA-GFP (gray bars) and AdK7-CA-GFP (black bars) determined by Sidak’s multiple comparisons test ( p

    Article Snippet: Isolated islets were cultured overnight in Roswell Park Memorial Institute (RPMI)-1640 medium (Sigma-Aldrich) containing 10% FBS (Nichirei Biosciences, Tokyo, Japan) and 5.5 mM glucose at 37°C and were subsequently treated with trypsin–ethylenediaminetetraacetic acid (EDTA) (Invitrogen, Carlsbad, CA, USA) for dispersion into single islet cells ( ).

    Techniques: Transduction, Expressing, Infection, Cell Culture, Flow Cytometry, Cytometry, Fluorescence, Standard Deviation

    Migration activity of EMT-induced cancer cells. (A and B) To measure cell migration activity, A549 (left panels) and Panc-1 (right panels) cells were subjected to the in vitro wound healing assay in the presence or absence (Control) of TGF-ß, as described in the text. Phase-contrast micrographs of the cultures were taken after 16-h incubation. Black broken lines indicate initial wound edges. Scale bar, 50 µm. B) Cell migration area was estimated by analyzing the pixel with Image J. Each bar indicates the mean ± SD for the areas of migrated cells in 3 different fields (right panels). (C) Cells that had been pretreated without or with TGF-ß for 24 h were incubated in 1% FBS-containing medium without or with TGF-ß onto a 24-well plate for 6 h at 37°C. The cell migration was monitored with a time-lapse video system for 12 h. Cell migration distance was measured for 15 randomly selected cells. Each bar indicates the mean ± SD for cell migration velocity of 15 cells. Other experimental conditions are described in Materials and Methods .

    Journal: PLoS ONE

    Article Title: Epithelial-Mesenchymal Transition Stimulates Human Cancer Cells to Extend Microtubule-based Invasive Protrusions and Suppresses Cell Growth in Collagen Gel

    doi: 10.1371/journal.pone.0053209

    Figure Lengend Snippet: Migration activity of EMT-induced cancer cells. (A and B) To measure cell migration activity, A549 (left panels) and Panc-1 (right panels) cells were subjected to the in vitro wound healing assay in the presence or absence (Control) of TGF-ß, as described in the text. Phase-contrast micrographs of the cultures were taken after 16-h incubation. Black broken lines indicate initial wound edges. Scale bar, 50 µm. B) Cell migration area was estimated by analyzing the pixel with Image J. Each bar indicates the mean ± SD for the areas of migrated cells in 3 different fields (right panels). (C) Cells that had been pretreated without or with TGF-ß for 24 h were incubated in 1% FBS-containing medium without or with TGF-ß onto a 24-well plate for 6 h at 37°C. The cell migration was monitored with a time-lapse video system for 12 h. Cell migration distance was measured for 15 randomly selected cells. Each bar indicates the mean ± SD for cell migration velocity of 15 cells. Other experimental conditions are described in Materials and Methods .

    Article Snippet: These cell lines were cultured in DMEM/F12 medium (Invitrogen, Carlsbed, CA) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences, Tokyo) at 37°C in a humidified atmosphere of 5% CO2 and 95% air.

    Techniques: Migration, Activity Assay, In Vitro, Wound Healing Assay, Incubation

    Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern

    doi: 10.1292/jvms.18-0202

    Figure Lengend Snippet: Photomicrographs of monolayer cultured canine bone marrow-derived mesenchymal stem cells (cBMSC) maintained in 10% FBS DMEM. (A) Day 1, primary culture (P0) showing non-adherent mononuclear cells immediately following seeding, (B) Day 4, P0 culture showing appearance of adherent elongated spindle-shaped fibroblastic-like cells just before removing non-adherent cells, (C) Day 10, P0 culture showing circular patches of small bipolar colony forming unit-fibroblasts (CFU-F), (D) Day 4, first passage (P1) culture showing bipolar to polygonal shaped cells, (E) Day 5, second passage (P2) and (f) Day 6, third passage (P3) showing larger polygonal shaped cells with reduced CFU-F formation. Scale Bars: 50 µ m.

    Article Snippet: After centrifugation, MC were collected from the sample/medium interface and plated in polystyrene culture plates containing Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO BRL, Grand Island, NY, U.S.A.) supplemented with 10% fetal bovine serum (FBS) (Nichirei Biosciences Inc., Tokyo, Japan, Batch #: 83300104), 100 IU/ml of penicillin and 0.1 mg/ml of streptomycin and incubated at 37°C in 5% CO2 in a humidified chamber without disturbing the plates for 4 days.

    Techniques: Cell Culture, Derivative Assay

    Recapitulation of FOP phenotype shown by two chondrogenesis pathways. (A) Schematic view of a protocol for MSC-like cell induction. Induced SM were treated with FGF2 (4 ng/ml) and FBS (10%) for 12 days. (B) FACS analysis using surface markers for MSCs at day 12 of MSC-like cell induction. CD44 + , CD73 + , CD105 + and CD45 (PTPRC) − cells were induced. SM cells were used as control populations. (C) Schematic view of MSC-like cell-CI and SCL-CI using FOP-iPSCs and resFOP-iPSCs. Briefly, induced MSC-like cells and SCL were spotted onto fibronectin-coated dishes and treated with CI basal media supplemented with activin A (30 ng/ml) for 5 days. (D) ACVR1 expression in MSC-like cells and SCL induced from both FOP-iPSCs and resFOP-iPSCs was assessed by RT-qPCR. Error bars represent s.e.m. ( n =6). The expression level of resFOP-MSCs was set to 1. (E-G) Evaluation of MSC-like cell-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (E), Alcian Blue staining (F) and GAG/DNA analysis (G). (H-J) Evaluation of SCL-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (H), Alcian Blue staining (I) and GAG/DNA analysis (J). (K,L) Evaluation of R667 and Rapamycin efficacy on MSC-like cell-CI. Chondrogenic differentiation was assessed at day 5 of CI by Alcian Blue staining (K) and GAG/DNA analysis (L). (M-P) In vitro study regarding cell-of-origins of the ectopic bone observed in FOP. PDGFRα + /CD31 – and PDGFRα – /CD31 – populations were isolated by FACS (M). Chondrogenic potential of each population was assessed at day 5 of CI by GAG/DNA (N) and RT-qPCR analysis (O). Expression levels of PAI1 and MMP1 , both surrogate markers of aberrant FOP-ACVR1 signaling, were higher in PDGFRα + /CD31 – cells (P). Error bars represent s.e.m. ( n =3). * P

    Journal: Development (Cambridge, England)

    Article Title: Modeling human somite development and fibrodysplasia ossificans progressiva with induced pluripotent stem cells

    doi: 10.1242/dev.165431

    Figure Lengend Snippet: Recapitulation of FOP phenotype shown by two chondrogenesis pathways. (A) Schematic view of a protocol for MSC-like cell induction. Induced SM were treated with FGF2 (4 ng/ml) and FBS (10%) for 12 days. (B) FACS analysis using surface markers for MSCs at day 12 of MSC-like cell induction. CD44 + , CD73 + , CD105 + and CD45 (PTPRC) − cells were induced. SM cells were used as control populations. (C) Schematic view of MSC-like cell-CI and SCL-CI using FOP-iPSCs and resFOP-iPSCs. Briefly, induced MSC-like cells and SCL were spotted onto fibronectin-coated dishes and treated with CI basal media supplemented with activin A (30 ng/ml) for 5 days. (D) ACVR1 expression in MSC-like cells and SCL induced from both FOP-iPSCs and resFOP-iPSCs was assessed by RT-qPCR. Error bars represent s.e.m. ( n =6). The expression level of resFOP-MSCs was set to 1. (E-G) Evaluation of MSC-like cell-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (E), Alcian Blue staining (F) and GAG/DNA analysis (G). (H-J) Evaluation of SCL-CI using FOP-iPSCs and resFOP-iPSCs. Chondrogenic differentiation was assessed at day 5 of CI by RT-qPCR analysis (H), Alcian Blue staining (I) and GAG/DNA analysis (J). (K,L) Evaluation of R667 and Rapamycin efficacy on MSC-like cell-CI. Chondrogenic differentiation was assessed at day 5 of CI by Alcian Blue staining (K) and GAG/DNA analysis (L). (M-P) In vitro study regarding cell-of-origins of the ectopic bone observed in FOP. PDGFRα + /CD31 – and PDGFRα – /CD31 – populations were isolated by FACS (M). Chondrogenic potential of each population was assessed at day 5 of CI by GAG/DNA (N) and RT-qPCR analysis (O). Expression levels of PAI1 and MMP1 , both surrogate markers of aberrant FOP-ACVR1 signaling, were higher in PDGFRα + /CD31 – cells (P). Error bars represent s.e.m. ( n =3). * P

    Article Snippet: MSC-like cell induction was carried out by replacing SM induction medium with MSC-like cell induction medium [10% FBS (Nichirei) and 4 ng/ml FGF2 (Wako) in αMEM (Nacalai Tesque)].

    Techniques: FACS, Expressing, Quantitative RT-PCR, Staining, In Vitro, Isolation