Human Umbilical Cord Blood-Derived Serum for Culturing the Supportive Feeder Cells of Human Pluripotent Stem Cell Lines

Figure 4

Characterization of human pluripotent stem cells after coculture with HFF-hUCS feeder cells. Two karyotypic normal hPSC lines, hESC (Chula2.hES) and hiPSC (HFSK#11.hiPS), were cocultured with inactivated HFF-hUCS or inactivated HFF-FBS, and subsequently the morphological appearance, pluripotent gene expression, and pluripotent marker expression were examined. hPSC lines showed undifferentiated colonies with defined boundaries and distinct prominent nuclei (a). hPSCs cocultured with inactivated HFF-hUCS expressed pluripotent genes, including OCT-4, NANOG, REX1, and UTF, similar to hPSCs cocultured with inactivated HFF-FBS (b). The pluripotent markers, including SSEA-3, TRA-1-60, TRA-1-81, and OCT-4, were detected in hPSCs cocultured with inactivated HFF-hUCS and inactivated HFF-FBS (c). The quantitative analysis of SSEA-4 expression through flow cytometry demonstrated that the percentage of SSEA-4-positive cells was not significantly different between hESCs cocultured with inactivated HFF-FBS and inactivated HFF-hUCS ( versus , resp.). The percentage of SSEA-4-positive hiPSCs cocultured with inactivated HFF-hUCS () was significantly higher than that for hiPSCs cocultured with inactivated HFF-FBS () (; (d)). HFF = human foreskin fibroblasts, FBS = fetal bovine serum, hUCS = human cord blood-derived serum, and DAPI = 4′,6-diamidino-2-phenylindole. Scale bars = 200 μm.

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