Journal: Stem Cell Research & Therapy

Article Title: Development of a surrogate potency assay to determine the angiogenic activity of Stempeucel®, a pooled, ex-vivo expanded, allogeneic human bone marrow mesenchymal stromal cell product

doi: 10.1186/s13287-017-0488-3

Figure Lengend Snippet: a qRT-PCR analysis of seven angiogenic genes: VEGF, HGF, SDF-1, Ang-1, IL-6, IL-8 and TGF-β1 in six batches of phBMMSCs. Relative fold expression was depicted after normalization to the housekeeping gene β-Actin as an internal standard and subsequent normalization using human foreskin fibroblast ( HFF ) cells for the same genes. b Secretome analysis of control medium and phBMMSC CM using 41 growth factor antibody arrays detected by chemiluminescence; a few detected growth factors marked in boxes . c Quantification of seven selected angiogenic growth factors/cytokines using human-specific ELISAs in same six batches of phBMMSCs. d Quantification of VEGF by ELISA in CM from five batches of phBMMSCs from P4 to P7; one-way ANOVA revealed no significant difference in VEGF levels between P4 and P7, p = 0.699. Data presented as mean ± SEM. SEMs shown as error bars. phBMMSC pooled human bone marrow derived mesenchymal stromal cell, Ang-1 angiopoietin-1, SDF-1 stromal-derived factor-1, bFGF basic fibroblast growth factor, HGF hepatocyte growth factor, IL interleukin, TGFβ1 transforming growth factor beta-1, VEGF vascular endothelial growth factor

Article Snippet: HUVECs were allowed to proliferate for 72 h. Then 5-bromo-2′-deoxyuridine (100 μM) was added (1:1000 in SFM) at the 48th hour, and its uptake by the cells was measured using a colorimetric cell proliferation ELISA kit (Merck-Millipore, NJ, USA).

Techniques: Quantitative RT-PCR, Expressing, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Development of a surrogate potency assay to determine the angiogenic activity of Stempeucel®, a pooled, ex-vivo expanded, allogeneic human bone marrow mesenchymal stromal cell product

doi: 10.1186/s13287-017-0488-3

Figure Lengend Snippet: Quantification and validation of VEGF secreted by phBMMSCs. a VEGF levels in the CM from twenty six batches of phBMMSCs by ELISA. VEGF level at 48 h was 1.7 ± 0.7 ng/ml/million cells and at 72 h was 2.8 ± 1.0 ng/ml/million cells. b Quantification of VEGF levels in the CM from ten batches of phBMMSCs estimated by two different operators; no significant differences were observed in the VEGF levels estimated by two different operators determined by Student’s t test, p = 0.774. c Quantification of VEGF levels in the CM from ten batches of phBMMSCs estimated by a single operator on two different days; no significant differences were observed in the VEGF levels estimated by a single operator at different time points as determined by Student’s t test, p = 0.631. Data presented as mean ± SEM. SEMs shown as error bars. VEGF vascular endothelial growth factor

Article Snippet: HUVECs were allowed to proliferate for 72 h. Then 5-bromo-2′-deoxyuridine (100 μM) was added (1:1000 in SFM) at the 48th hour, and its uptake by the cells was measured using a colorimetric cell proliferation ELISA kit (Merck-Millipore, NJ, USA).

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay