Enhanced Generation of Human Induced Pluripotent Stem Cells from Peripheral Blood and Using Their Mesoderm Differentiation Ability to Regenerate Infarcted Myocardium

Figure 2

Mesoderm differentiation preference in the process of CiMS-iPSC differentiation to three germ layers. (a) Expression of pluripotent markers on day 0 and day 2 of mesodermal differentiation was estimated by real-time PCR. The expression of early mesodermal markers on day 2 of mesoderm differentiation was compared in each cell type using real-time PCR. The expression of early endodermal markers on day 4 of endodermal differentiation and early ectodermal markers on day 5 of ectodermal differentiation was compared in each cell type using real-time PCR (, , and ; ss: statistically significant; ns: statistically not significant; black bar: CiMS-iPSC; white bar: HDF-iPSC; grey bar: hESC). (b) After differentiation into cardiovascular lineage cells, the expression of endothelial cell (EC) markers (CD34, KDR, and CD31) on day 5 of EC differentiation, vascular smooth muscle cell (VSMC) markers (SMA, SM22a, and CNN1) on day 6 of VSMC differentiation, and cardiomyocyte (CMC) markers (Nkx2.5, GATA4) on day 12 of CMC differentiation was compared in each cell type using real-time PCR. (c) After differentiation into cardiovascular cells, such as ECs, VSMCs, and CMCs, cardiovascular cell-specific markers were detected by immunofluorescent staining, including CD31 for ECs, SMA for VSMCs, and alpha-sarcomeric actin for CMCs. (d) Marker-positive cells were calculated among the total differentiated cells, and differentiation efficiency was compared.

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