Fig. 5
ECM-adherent UCB-NHSCs express ES cell markers. a The gene expression profile of P2 umbilical cord blood-derived non-hematopoietic stem cells (UCB-NHSCs) cultured on ECM was determined using TaqMan PCR. RNA isolated from human embryonic stem (ES) (ESI6) cells was used as a positive control, while RNA harvested from human bone marrow-derived mesenchymal cells (hBM-MSCs) was used for normalization of the data. All three types of cells were prepared, cultured on ECM, and harvested in parallel. The data shown represent the mean ± SD fold-changes (note: log base 10 scale on y axis) in transcript level after normalization to that of BM-MSCs. b Immunofluorescent (IF) detection of OCT4, SSEA-4, and TRA-1-60. UCB-NHSCs (P2) pre-cultured on ECM were seeded (2.5–7.5 × 104 cells/cm2) onto regular chamber slides and cultured for 2 days. Cells were immunostained with antibodies specific for the proteins of interest. Non-specific isotype rabbit IgG, mouse IgG, and mouse IgM were used as negative controls (Neg. cont.) for OCT4, SSEA-4, and TRA-1-60, respectively. Parallel cultures of BM-MSCs were also prepared and treated similarly. Phase contrast micrographs of both types of cells are shown for comparative purposes. Scale bar = 10 μm