Figure. 4.

Enhanced cytokine expression by ESHF derived CDCs. Expression levels of several key cytokines were determined by RT-PCR after 16 hours of hypoxia. () ESHF derived CDCs showed up regulated expression of VEGF-1A (*P=0.02), SDF-1α (*P=0.019) and HIF-1α (*P=0.032). (D–I) Expression for IGF1, IL-8, PGDFB, bFGF, CXCL1, and OSGIN1 did not show any significant difference between CHD and ESHF derived CDCs. To determine if the augmentation of pro-angiogenic factors in ESHF derived CDCs translated to increased vessel formation, in vitro endothelial cell tube-formation assay was performed using human umbilical vein endothelial cells (HUVEC). (J) Representative images of HUVECs 8 hours after plating on matrix-coated wells in serum free basal medium (IMDM), HUVEC condition media, CHD derived CDCs condition medium and ESHF derived CDCs condition medium and cardio-fibroblast condition medium (Cfb). Quantification of total tube length demonstrated significantly increased tube length with ESHF derived CDCs as compared to CHD derived CDCs, IMDM and Cfb (scale bar = 140µm). (**=P<0.01 and ***=P<0.001 (K–M) Quantitative rt-PCR analysis of myocardial biopsies obtained from CHD or ESHF patients demonstrated increased HSF-1 (*P<0.05), HSP 60 (*P<0.05), and HSP 70 (*P<0.05) in ESHF myocardium. Data are analyzed one-Way ANOVA followed by Dunns analysis or non-parametric t-test followed by Mann-Whitney’s analysis and presented as mean ± SEM.