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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: 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

Fig. 5

In vitro pro-angiogenic function of secreted VEGF from nine batches of phBMMSCs. a Evaluation of HUVEC migration under various culture conditions. CM (n = 9) induced significant migration of HUVECs. Migration was unaffected by an isotype-matched antibody (p = 0.1964), while addition of anti-VEGF mAb caused significant (*p = 0.0001) inhibition of cell migration. Migration of HUVECs in the CM group was significantly higher than that of control medium, ***p = 0.0004. b Correlation of VEGF concentration in phBMMSC CM (n = 9) with HUVEC migration was observed, r 2 = 0.78, **p = 0.0014. c HUVEC proliferation in response to various culture conditions; phBMMSC CM (n = 9) induced HUVEC proliferation similar to that of EGM and complete medium, although significant reduction of cell proliferation was observed in the CM group with anti-VEGF mAb in comparison to CM with isotype IgG, **p = 0.0075. Addition of IgG isotype in the CM did not alter HUVEC proliferation compared with CM alone, p = 0.2985. d Correlation of VEGF concentration-dependent proliferation of HUVECs was observed with phBMMSC CM (n = 9) with a high correlation coefficient, r 2 = 0.75, **p = 0.002. Representative images of tube formations in e EGM, f control medium, g phBMMSC CM, h IgG isotype, i phBMMSC CM with anti-VEGF mAb, 4× objective were used for magnification. j phBMMSC CM (n = 9) induces significant tube formation in HUVECs compared with EGM and control medium (**p = 0.0063). Tube formation was greatly reduced with the addition of anti-VEGF mAb in the CM in comparison with the isotype-matched control group (**p = 0.0084). Tube formation in HUVECs was unaffected with the addition of IgG isotype (p = 0.2985). k Concentration of VEGF in nine batches of CM highly correlated to the tube lengths, r 2 = 0.89, ***p = 0.0001. Data presented as mean ± SEM. SEMs shown as error bars. CM conditioned medium, phBMMSC pooled human bone marrow derived mesenchymal stromal cell, EGM endothelial growth medium, VEGF vascular endothelial growth factor, HPF high-power field

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