Fig. 2

Characterization of induced pluripotent stem cell (iPSC) derived from fibroblastic and epithelial cells and their functional properties. (a) Detection of pluripotency-associated markers. The representative images of human iPSC colonies in established lines show a typical embryonic stem cell-like morphology and demonstration of alkaline phosphatase (AP) activity in studied cells. iPSCs derived from neonatal and adult fibroblasts as well as epithelial cells were positive for NANOG, OCT3/4, and C-MYC transcription factors and displayed TRA-1-60 and TRA-1-81 expression on their surface. Human embryonic stem cell (hESC) line BG01V was used as a positive control. Shown are representative immunofluorescence images captured by using an Eclipse Ci-S epifluorescence microscope with 20× objective. Scale bar = 100 μm. (b) Embryoid body (EB)-mediated multilineage differentiation of episomally generated iPSC lines. Phase-contrast images showing the EBs formed from studied iPSC lines. EBs were spontaneously differentiated and examined for the presence of markers of the three germ layers. Immunostainings show cells positive for ectodermal marker MAP2, the mesodermal marker alpha-smooth muscle actin (αSMA), and the endodermal marker SOX17. The trilineage potential is additionally evidenced by the presence of cells with morphology characteristic for neurons and cytoskeleton typical for mesodermal cells. Scale bar = 100 μm. AmiPSC amniocyte-derived induced pluripotent stem cell, BJiPSC induced pluripotent stem cell derived from BJ cell, StiPSC induced pluripotent stem cell derived from scar tissue fibroblast, UiPSC induced pluripotent stem cell reprogrammed from urinary epithelial cells