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    Name:
    Human Mesenchymal Stem Cell Adipogenic Differentiation Medium BulletKitTM
    Description:
    Culture system containing basal medium and supplements required for maintenance and induction of hMSC adipogenic differentiation
    Catalog Number:
    PT-3004
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    Category:
    Primary and Stem Cells
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    Structured Review

    Lonza hbm mscs hbm mscs
    Effects of endocannabinoids on adipogenesis in <t>hBM-MSCs.</t> hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects of endocannabinoids were evaluated in hBM-MSCs. NADA (A), AEA (B) and 2-AG (C), were treated for 72 hours in hBM-MSC culture. The cell viability was determined with a detecting reagent, 10 μM of WST-1. Adipogenesis was induced in hBM-MSCs under the presence of the IDX adipocyte differentiation inducing medium. After treating NADA, AEA and 2-AG, in every two or three day, media were exchanged. At the 7 th days in culture, lipid droplets in differentiated adipocytes were stained with Oil Red O (ORO) (D). After dissolving the ORO in isopropyl alcohol, the level of staining was quantified at 500 nm using a spectrometer. Data were normalized by setting the control as 1 (E). Results are the mean ± standard deviation (SD) of three measurements using independent hBM-MSCs from three different donors (n=3). * p ≤0.05 and ** p ≤0.01.
    Culture system containing basal medium and supplements required for maintenance and induction of hMSC adipogenic differentiation
    https://www.bioz.com/result/hbm mscs hbm mscs/product/Lonza
    Average 96 stars, based on 1 article reviews
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    hbm mscs hbm mscs - by Bioz Stars, 2021-06
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    1) Product Images from "A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells"

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    Journal: Biomolecules & Therapeutics

    doi: 10.4062/biomolther.2014.137

    Effects of endocannabinoids on adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects of endocannabinoids were evaluated in hBM-MSCs. NADA (A), AEA (B) and 2-AG (C), were treated for 72 hours in hBM-MSC culture. The cell viability was determined with a detecting reagent, 10 μM of WST-1. Adipogenesis was induced in hBM-MSCs under the presence of the IDX adipocyte differentiation inducing medium. After treating NADA, AEA and 2-AG, in every two or three day, media were exchanged. At the 7 th days in culture, lipid droplets in differentiated adipocytes were stained with Oil Red O (ORO) (D). After dissolving the ORO in isopropyl alcohol, the level of staining was quantified at 500 nm using a spectrometer. Data were normalized by setting the control as 1 (E). Results are the mean ± standard deviation (SD) of three measurements using independent hBM-MSCs from three different donors (n=3). * p ≤0.05 and ** p ≤0.01.
    Figure Legend Snippet: Effects of endocannabinoids on adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects of endocannabinoids were evaluated in hBM-MSCs. NADA (A), AEA (B) and 2-AG (C), were treated for 72 hours in hBM-MSC culture. The cell viability was determined with a detecting reagent, 10 μM of WST-1. Adipogenesis was induced in hBM-MSCs under the presence of the IDX adipocyte differentiation inducing medium. After treating NADA, AEA and 2-AG, in every two or three day, media were exchanged. At the 7 th days in culture, lipid droplets in differentiated adipocytes were stained with Oil Red O (ORO) (D). After dissolving the ORO in isopropyl alcohol, the level of staining was quantified at 500 nm using a spectrometer. Data were normalized by setting the control as 1 (E). Results are the mean ± standard deviation (SD) of three measurements using independent hBM-MSCs from three different donors (n=3). * p ≤0.05 and ** p ≤0.01.

    Techniques Used: Cell Culture, Staining, Standard Deviation

    Transcriptional expression profile of CNR1, CNR2 and TRPV1 during adipogenesis in hBM-MSCs. Adipocyte differentiation was induced in hBM-MSCs by exchanging culture media supplemented with 1 μg/ml insulin, 0.1 μM dexamethasone and 0.5 mM isobutylmethylxanthine (IDX). At the seventh day after the induction of adipogenesis, total RNA was extracted and Q-RT-PCR analysis was performed for (A) CNR1 and (B) TRPV1. Values represent the mean expression ± SE of the mRNA of the various genes relative to human GAPDH expression (n=3), * p ≤0.05, ** p ≤0.01.
    Figure Legend Snippet: Transcriptional expression profile of CNR1, CNR2 and TRPV1 during adipogenesis in hBM-MSCs. Adipocyte differentiation was induced in hBM-MSCs by exchanging culture media supplemented with 1 μg/ml insulin, 0.1 μM dexamethasone and 0.5 mM isobutylmethylxanthine (IDX). At the seventh day after the induction of adipogenesis, total RNA was extracted and Q-RT-PCR analysis was performed for (A) CNR1 and (B) TRPV1. Values represent the mean expression ± SE of the mRNA of the various genes relative to human GAPDH expression (n=3), * p ≤0.05, ** p ≤0.01.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    The effects of rimonabant on adipogenesis in hBM-MSCs and PPARγ transactivation. (A) The concentration-dependent effect of rimonabant on adipogenesis in hBM-MSCs was evaluated in 24 well plates. The effect of rimonabant was compared with those of aspirin, glibenclamide and troglitazone. Cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. (B) CV-1 cells were transiently cotransfected with the PPARγ expression vector and the PPRE-TK-Luciferase reporter, and then treated with vehicle, rimonabant (RIMO), AEA, NADA and 2-AG (1, 3, and 10 μM) or troglitazone (TRO, 1 μM). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.
    Figure Legend Snippet: The effects of rimonabant on adipogenesis in hBM-MSCs and PPARγ transactivation. (A) The concentration-dependent effect of rimonabant on adipogenesis in hBM-MSCs was evaluated in 24 well plates. The effect of rimonabant was compared with those of aspirin, glibenclamide and troglitazone. Cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. (B) CV-1 cells were transiently cotransfected with the PPARγ expression vector and the PPRE-TK-Luciferase reporter, and then treated with vehicle, rimonabant (RIMO), AEA, NADA and 2-AG (1, 3, and 10 μM) or troglitazone (TRO, 1 μM). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Techniques Used: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Plasmid Preparation, Luciferase, Standard Deviation

    Effects of endocannabinoids on adipocyte differentiation marker expression during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. At the 7 th days in culture, total RNA samples were extracted and Q-RT-PCR was performed for PPARγ (A), FABP4 (B) and adiponectin (C). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.
    Figure Legend Snippet: Effects of endocannabinoids on adipocyte differentiation marker expression during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. At the 7 th days in culture, total RNA samples were extracted and Q-RT-PCR was performed for PPARγ (A), FABP4 (B) and adiponectin (C). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Techniques Used: Marker, Expressing, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

    Effects of endocannabinoids on adiponectin production during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. ELISA was performed to measure the concentration of adiponectin (A) accumulated in cell culture supernatants for 48 hours after the last medium exchange. The concentration-dependent effect of arachidonyl dopamine (NADA) on the inhibition of adipogenesis was evaluated (B). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.
    Figure Legend Snippet: Effects of endocannabinoids on adiponectin production during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. ELISA was performed to measure the concentration of adiponectin (A) accumulated in cell culture supernatants for 48 hours after the last medium exchange. The concentration-dependent effect of arachidonyl dopamine (NADA) on the inhibition of adipogenesis was evaluated (B). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Concentration Assay, Cell Culture, Inhibition, Expressing, Standard Deviation

    Effects of rimonabant and capsazepine on the NADA-dependent inhibition of adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects were evaluated for rimonabant (A) and capsazepine (B), both of which were treated for 72 hours in hBM-MSC culture. The cell viability was determined by WST-1 assay. For testing the effects of capsazepine (C) and rimonabant (D) on the NADA-induced inhibition on the adipogenesis in hBM-MSCs, cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.
    Figure Legend Snippet: Effects of rimonabant and capsazepine on the NADA-dependent inhibition of adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects were evaluated for rimonabant (A) and capsazepine (B), both of which were treated for 72 hours in hBM-MSC culture. The cell viability was determined by WST-1 assay. For testing the effects of capsazepine (C) and rimonabant (D) on the NADA-induced inhibition on the adipogenesis in hBM-MSCs, cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Techniques Used: Inhibition, Cell Culture, WST-1 Assay, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation

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    Article Snippet: .. hBM-MSCs were purchased from Lonza (Walkersville, MD, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; glucose 1 g/L) containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA). ..

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    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells
    Article Snippet: .. Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA). .. To induce adipogenesis, hBM-MSCs were cultured to 100% confluence.

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    Modification:

    Article Title: Polypharmacology of N6-(3-Iodobenzyl)adenosine-5′-N-methyluronamide (IB-MECA) and Related A3 Adenosine Receptor Ligands: Peroxisome Proliferator Activated Receptor (PPAR) γ Partial Agonist and PPARδ Antagonist Activity Suggests Their Antidiabetic Potential
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    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells
    Article Snippet: .. Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA). .. To induce adipogenesis, hBM-MSCs were cultured to 100% confluence.

    Functional Assay:

    Article Title: Real-Time Monitoring of Glutathione in Living Cells Reveals that High Glutathione Levels Are Required to Maintain Stem Cell Function
    Article Snippet: The cells were grown in phenol red-free DMEM (Welgene, LM 001-10; Gyeongsan, Korea) supplemented with 10% heat-inactivated fetal bovine serum (HyClone; GE Healthcare, Melbourne, VIC, Australia), 100 U/mL penicillin, 100 μg/mL streptomycin sulfate, and 2 mM glutamine. hES-MSCs were kindly provided by Hyung-Min Chung (Konkuk University, Korea). .. Cellular GSH of hES-MSCs was depleted by treating cells with 80 μM BSO (Sigma-Aldrich, St. Louis, MO) for 24 hr and was restored with 1 mM GSH-EE (Sigma-Aldrich) for 2 hr, followed by their functional assays. hBM-MSCs (Lonza, Walkersville, MD) were cultured according to the manufacturer's instructions. .. Culture, EB formation, and in vitro neuronal differentiation of murine E14TG2a ESCs (ATCC) were performed as previously described ( ).

    Irradiation:

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow
    Article Snippet: .. The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S. ..

    Knock-Out:

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow
    Article Snippet: .. The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S. ..

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    Lonza hbm mscs
    Evaluation of adiponectin-promoting activity of 1a and related A 3 AR ligands. (A) 1a , 1b , 1c , or 3a was co-treated with the IDX medium in <t>hBM-MSCs.</t> On the 7th day in culture, cell culture supernatants were harvested and ELISA was performed to measure levels of adiponectin. (B) The effects of A 3 AR agonists 1a , 1c , and 17 and A 3 AR antagonist 18 on adiponectin production were evaluated. The pharmacological correlation between A 1 AR (C), A 2A AR (D), A 3 AR (E) binding K i value at 1 µM and adiponectin levels was analyzed. Values represent mean ± SD ( n = 3, three independent experiments): (*) p ≤ 0.05 and (**) p ≤ 0.01.
    Hbm Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hbm mscs/product/Lonza
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hbm mscs - by Bioz Stars, 2021-06
    96/100 stars
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    97
    Lonza primary human bone marrow derived mscs
    Long-term monitoring of human <t>MSCs</t> in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 <t>hBM-MSC</t> or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment
    Primary Human Bone Marrow Derived Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary human bone marrow derived mscs/product/Lonza
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary human bone marrow derived mscs - by Bioz Stars, 2021-06
    97/100 stars
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    Evaluation of adiponectin-promoting activity of 1a and related A 3 AR ligands. (A) 1a , 1b , 1c , or 3a was co-treated with the IDX medium in hBM-MSCs. On the 7th day in culture, cell culture supernatants were harvested and ELISA was performed to measure levels of adiponectin. (B) The effects of A 3 AR agonists 1a , 1c , and 17 and A 3 AR antagonist 18 on adiponectin production were evaluated. The pharmacological correlation between A 1 AR (C), A 2A AR (D), A 3 AR (E) binding K i value at 1 µM and adiponectin levels was analyzed. Values represent mean ± SD ( n = 3, three independent experiments): (*) p ≤ 0.05 and (**) p ≤ 0.01.

    Journal: Journal of medicinal chemistry

    Article Title: Polypharmacology of N6-(3-Iodobenzyl)adenosine-5′-N-methyluronamide (IB-MECA) and Related A3 Adenosine Receptor Ligands: Peroxisome Proliferator Activated Receptor (PPAR) γ Partial Agonist and PPARδ Antagonist Activity Suggests Their Antidiabetic Potential

    doi: 10.1021/acs.jmedchem.7b00805

    Figure Lengend Snippet: Evaluation of adiponectin-promoting activity of 1a and related A 3 AR ligands. (A) 1a , 1b , 1c , or 3a was co-treated with the IDX medium in hBM-MSCs. On the 7th day in culture, cell culture supernatants were harvested and ELISA was performed to measure levels of adiponectin. (B) The effects of A 3 AR agonists 1a , 1c , and 17 and A 3 AR antagonist 18 on adiponectin production were evaluated. The pharmacological correlation between A 1 AR (C), A 2A AR (D), A 3 AR (E) binding K i value at 1 µM and adiponectin levels was analyzed. Values represent mean ± SD ( n = 3, three independent experiments): (*) p ≤ 0.05 and (**) p ≤ 0.01.

    Article Snippet: hBM-MSCs were purchased from Lonza (Walkersville, MD, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; glucose 1 g/L) containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA).

    Techniques: Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Binding Assay

    Independency of A3 AR Signaling on 1a-Induced Upregulation of Adiponectin Production in hBM-MSCs

    Journal: Journal of medicinal chemistry

    Article Title: Polypharmacology of N6-(3-Iodobenzyl)adenosine-5′-N-methyluronamide (IB-MECA) and Related A3 Adenosine Receptor Ligands: Peroxisome Proliferator Activated Receptor (PPAR) γ Partial Agonist and PPARδ Antagonist Activity Suggests Their Antidiabetic Potential

    doi: 10.1021/acs.jmedchem.7b00805

    Figure Lengend Snippet: Independency of A3 AR Signaling on 1a-Induced Upregulation of Adiponectin Production in hBM-MSCs

    Article Snippet: hBM-MSCs were purchased from Lonza (Walkersville, MD, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; glucose 1 g/L) containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA).

    Techniques:

    Validation of 1a and related A 3 AR ligands as PPAR δ antagonists. (A) The effects of 1a , 1c , 3a , and 23 (1 µM) on the 22 (0.03 µM) induced interaction between fluorescein-C33 coactivator peptides and PPAR δ LBD in a TR-FRET PPAR δ coactivator assay were evaluated. (B) In a TR-FRET PPAR δ corepressor assay with SMRT-ID2 peptides, the activities of 23 , 24 , and 3a (1 µM) were assessed in the condition that 22 (0.1 µM) existed or not. (C) 23 , 24 , 3a , and 1c were evaluated in a TR-FRET PPAR δ corepressor assay with SMRT-ID2 peptides at various concentrations. For functional validation assay for PPAR δ antagonism, hBM-MSCs were differentiated in the IDX condition and co-treated with 23 , 24 , or 3a in the medium. On the 3rd day in culture, total RNA was extracted and Q-RT-PCR was performed for ANGPTL4 (D) and PDK4 (E). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean ± SD ( n = 3, three independent experiments): (*) p ≤ 0.05 and (**) p ≤ 0.01.

    Journal: Journal of medicinal chemistry

    Article Title: Polypharmacology of N6-(3-Iodobenzyl)adenosine-5′-N-methyluronamide (IB-MECA) and Related A3 Adenosine Receptor Ligands: Peroxisome Proliferator Activated Receptor (PPAR) γ Partial Agonist and PPARδ Antagonist Activity Suggests Their Antidiabetic Potential

    doi: 10.1021/acs.jmedchem.7b00805

    Figure Lengend Snippet: Validation of 1a and related A 3 AR ligands as PPAR δ antagonists. (A) The effects of 1a , 1c , 3a , and 23 (1 µM) on the 22 (0.03 µM) induced interaction between fluorescein-C33 coactivator peptides and PPAR δ LBD in a TR-FRET PPAR δ coactivator assay were evaluated. (B) In a TR-FRET PPAR δ corepressor assay with SMRT-ID2 peptides, the activities of 23 , 24 , and 3a (1 µM) were assessed in the condition that 22 (0.1 µM) existed or not. (C) 23 , 24 , 3a , and 1c were evaluated in a TR-FRET PPAR δ corepressor assay with SMRT-ID2 peptides at various concentrations. For functional validation assay for PPAR δ antagonism, hBM-MSCs were differentiated in the IDX condition and co-treated with 23 , 24 , or 3a in the medium. On the 3rd day in culture, total RNA was extracted and Q-RT-PCR was performed for ANGPTL4 (D) and PDK4 (E). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean ± SD ( n = 3, three independent experiments): (*) p ≤ 0.05 and (**) p ≤ 0.01.

    Article Snippet: hBM-MSCs were purchased from Lonza (Walkersville, MD, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; glucose 1 g/L) containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA).

    Techniques: Functional Assay, Reverse Transcription Polymerase Chain Reaction

    Effects of endocannabinoids on adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects of endocannabinoids were evaluated in hBM-MSCs. NADA (A), AEA (B) and 2-AG (C), were treated for 72 hours in hBM-MSC culture. The cell viability was determined with a detecting reagent, 10 μM of WST-1. Adipogenesis was induced in hBM-MSCs under the presence of the IDX adipocyte differentiation inducing medium. After treating NADA, AEA and 2-AG, in every two or three day, media were exchanged. At the 7 th days in culture, lipid droplets in differentiated adipocytes were stained with Oil Red O (ORO) (D). After dissolving the ORO in isopropyl alcohol, the level of staining was quantified at 500 nm using a spectrometer. Data were normalized by setting the control as 1 (E). Results are the mean ± standard deviation (SD) of three measurements using independent hBM-MSCs from three different donors (n=3). * p ≤0.05 and ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: Effects of endocannabinoids on adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects of endocannabinoids were evaluated in hBM-MSCs. NADA (A), AEA (B) and 2-AG (C), were treated for 72 hours in hBM-MSC culture. The cell viability was determined with a detecting reagent, 10 μM of WST-1. Adipogenesis was induced in hBM-MSCs under the presence of the IDX adipocyte differentiation inducing medium. After treating NADA, AEA and 2-AG, in every two or three day, media were exchanged. At the 7 th days in culture, lipid droplets in differentiated adipocytes were stained with Oil Red O (ORO) (D). After dissolving the ORO in isopropyl alcohol, the level of staining was quantified at 500 nm using a spectrometer. Data were normalized by setting the control as 1 (E). Results are the mean ± standard deviation (SD) of three measurements using independent hBM-MSCs from three different donors (n=3). * p ≤0.05 and ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Cell Culture, Staining, Standard Deviation

    Transcriptional expression profile of CNR1, CNR2 and TRPV1 during adipogenesis in hBM-MSCs. Adipocyte differentiation was induced in hBM-MSCs by exchanging culture media supplemented with 1 μg/ml insulin, 0.1 μM dexamethasone and 0.5 mM isobutylmethylxanthine (IDX). At the seventh day after the induction of adipogenesis, total RNA was extracted and Q-RT-PCR analysis was performed for (A) CNR1 and (B) TRPV1. Values represent the mean expression ± SE of the mRNA of the various genes relative to human GAPDH expression (n=3), * p ≤0.05, ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: Transcriptional expression profile of CNR1, CNR2 and TRPV1 during adipogenesis in hBM-MSCs. Adipocyte differentiation was induced in hBM-MSCs by exchanging culture media supplemented with 1 μg/ml insulin, 0.1 μM dexamethasone and 0.5 mM isobutylmethylxanthine (IDX). At the seventh day after the induction of adipogenesis, total RNA was extracted and Q-RT-PCR analysis was performed for (A) CNR1 and (B) TRPV1. Values represent the mean expression ± SE of the mRNA of the various genes relative to human GAPDH expression (n=3), * p ≤0.05, ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    The effects of rimonabant on adipogenesis in hBM-MSCs and PPARγ transactivation. (A) The concentration-dependent effect of rimonabant on adipogenesis in hBM-MSCs was evaluated in 24 well plates. The effect of rimonabant was compared with those of aspirin, glibenclamide and troglitazone. Cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. (B) CV-1 cells were transiently cotransfected with the PPARγ expression vector and the PPRE-TK-Luciferase reporter, and then treated with vehicle, rimonabant (RIMO), AEA, NADA and 2-AG (1, 3, and 10 μM) or troglitazone (TRO, 1 μM). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: The effects of rimonabant on adipogenesis in hBM-MSCs and PPARγ transactivation. (A) The concentration-dependent effect of rimonabant on adipogenesis in hBM-MSCs was evaluated in 24 well plates. The effect of rimonabant was compared with those of aspirin, glibenclamide and troglitazone. Cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. (B) CV-1 cells were transiently cotransfected with the PPARγ expression vector and the PPRE-TK-Luciferase reporter, and then treated with vehicle, rimonabant (RIMO), AEA, NADA and 2-AG (1, 3, and 10 μM) or troglitazone (TRO, 1 μM). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Plasmid Preparation, Luciferase, Standard Deviation

    Effects of endocannabinoids on adipocyte differentiation marker expression during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. At the 7 th days in culture, total RNA samples were extracted and Q-RT-PCR was performed for PPARγ (A), FABP4 (B) and adiponectin (C). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: Effects of endocannabinoids on adipocyte differentiation marker expression during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. At the 7 th days in culture, total RNA samples were extracted and Q-RT-PCR was performed for PPARγ (A), FABP4 (B) and adiponectin (C). GAPDH was used as an internal control for Q-RT-PCR standardization. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Marker, Expressing, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

    Effects of endocannabinoids on adiponectin production during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. ELISA was performed to measure the concentration of adiponectin (A) accumulated in cell culture supernatants for 48 hours after the last medium exchange. The concentration-dependent effect of arachidonyl dopamine (NADA) on the inhibition of adipogenesis was evaluated (B). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: Effects of endocannabinoids on adiponectin production during adipogenesis in hBM-MSCs. When hBM-MSCs were in confluent state, adipogenesis was induced by exchanging media with the IDX adipogenic cocktail. ELISA was performed to measure the concentration of adiponectin (A) accumulated in cell culture supernatants for 48 hours after the last medium exchange. The concentration-dependent effect of arachidonyl dopamine (NADA) on the inhibition of adipogenesis was evaluated (B). Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Cell Culture, Inhibition, Expressing, Standard Deviation

    Effects of rimonabant and capsazepine on the NADA-dependent inhibition of adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects were evaluated for rimonabant (A) and capsazepine (B), both of which were treated for 72 hours in hBM-MSC culture. The cell viability was determined by WST-1 assay. For testing the effects of capsazepine (C) and rimonabant (D) on the NADA-induced inhibition on the adipogenesis in hBM-MSCs, cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Journal: Biomolecules & Therapeutics

    Article Title: A Cannabinoid Receptor Agonist N-Arachidonoyl Dopamine Inhibits Adipocyte Differentiation in Human Mesenchymal Stem Cells

    doi: 10.4062/biomolther.2014.137

    Figure Lengend Snippet: Effects of rimonabant and capsazepine on the NADA-dependent inhibition of adipogenesis in hBM-MSCs. hBM-MSCs were cultured in 24 well plates. When confluent, the cell viability effects were evaluated for rimonabant (A) and capsazepine (B), both of which were treated for 72 hours in hBM-MSC culture. The cell viability was determined by WST-1 assay. For testing the effects of capsazepine (C) and rimonabant (D) on the NADA-induced inhibition on the adipogenesis in hBM-MSCs, cell culture supernatants were harvested for the measurement of adiponectin by ELISA at the 7 th days after exchanging media containing with IDX. Values represent the mean expression ± standard deviation (SD) (n=3). * p ≤0.05 and ** p ≤0.01.

    Article Snippet: Cell culture and differentiation of hBM-MSCs hBM-MSCs were purchased from Lonza, Inc. (Walkersville, MD, USA). hBM-MSCs were cultured according to the manufacturer’s instructions with minor modifications. hBM-MSCs were maintained in Dulbecco’s modified eagle’s medium (DMEM) with low glucose (1 g/L glucose) containing 10% fetal bovine serum (FBS) (Lonza), supplemented with antibiotics and GlutamaxTM (Invitrogen, Carlsbad, CA, USA).

    Techniques: Inhibition, Cell Culture, WST-1 Assay, Enzyme-linked Immunosorbent Assay, Expressing, Standard Deviation

    Comparison of the fracture healing response by hBM-MSCs cultured in the (A) absence or (B) presence of osteogenic supplements, as shown by x-ray and μCT scans taken 8 wks following surgery.

    Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow

    doi: 10.1002/jor.21480

    Figure Lengend Snippet: Comparison of the fracture healing response by hBM-MSCs cultured in the (A) absence or (B) presence of osteogenic supplements, as shown by x-ray and μCT scans taken 8 wks following surgery.

    Article Snippet: The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S.

    Techniques: Cell Culture

    (A) Maximum torque and (B) Stiffness based on biomechanical testing of fractured femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow

    doi: 10.1002/jor.21480

    Figure Lengend Snippet: (A) Maximum torque and (B) Stiffness based on biomechanical testing of fractured femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Article Snippet: The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S.

    Techniques: Derivative Assay

    Fracture healing grades in the rat femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow

    doi: 10.1002/jor.21480

    Figure Lengend Snippet: Fracture healing grades in the rat femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Article Snippet: The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S.

    Techniques: Derivative Assay

    Comparative radiographic assessment of fracture healing in rat femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

    Article Title: Induction of Fracture Repair by Mesenchymal Cells Derived from Human Embryonic Stem Cells or Bone Marrow

    doi: 10.1002/jor.21480

    Figure Lengend Snippet: Comparative radiographic assessment of fracture healing in rat femurs that received no cells, CD73+ hESC-derived MSCs, or hBM-MSCs differentiated under similar osteogenic conditions for 7 days.

    Article Snippet: The hESC line H9 (obtained from WiCell) was maintained as undifferentiated cells as previously described ( , ) by coculture with irradiated mouse embryonic fibroblast (MEF) cells in DMEM/F12 supplemented with 15% Knockout Serum Replacer (KOSR) (Invitrogen), 1% MEM nonessential amino acids (Invitrogen), 0.5% penicillin-streptomycin (P/S), 2 mM L-Glutamine, 0.1 mM β-mercaptoethanol (Sigma), and 4 ng/mL human bFGF (Invitrogen). hBM-MSCs (Lonza) were cultured in phenol red-free αMEM (Invitrogen) supplemented with 10% FBS and 1% P/S.

    Techniques: Derivative Assay

    Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Journal: Stem Cell Research & Therapy

    Article Title: Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration

    doi: 10.1186/s13287-018-1076-x

    Figure Lengend Snippet: Long-term monitoring of human MSCs in BALB/c mice. a Representative BLI of mice administered with 5 × 10 5 hBM-MSC or hUC-MSC via the IC or IV route. The signal was progressively lost shortly after administration, with no evidence of malignant growth. b Ex vivo bioluminescence imaging of organs within 5 h of administration of the cells. Organs are indicated as the kidneys (k), spleen (s), liver (li), lungs (lu), heart (h) or brain (b). In some occasions, signal foci were seen in the heart of mice that received hUC-MSC IV (red arrow). c BLI images from mice that displayed hUC-MSC signal that persisted beyond day 7 after IV administration (ventral orientation, lower scale). In all cases, the signals had disappeared by day 21 and had not returned by the end of the experiment

    Article Snippet: Cell preparation Mouse kidney-derived stem cells (mKSCs) [ ], the D1 mouse MSC (mMSC) line (D1 ORL UVA [D1](ATCC® CRL-12424™)), primary human umbilical cord-derived MSCs (hUC-MSCs; collected from consenting donors and produced identically to those already being used in clinical trials by NHS Blood and Transplant (NHSBT)), primary human bone marrow-derived MSCs (hBM-MSCs; Lonza PT-2501), human kidney cells (hKCs; the kidneys deemed unsuitable for transplantation via UK NHSBT [ ]) and RAW264.7 macrophages (European Collection of Authenticated Cell Cultures 91062702) were cultured at 37 °C under a humidified atmosphere with 5% CO2 (culture media are described in Additional file ).

    Techniques: Mouse Assay, Ex Vivo, Imaging