Fig. 1
Newly isolated SSCs transformed into pluripotent state during long-term culture. a-d Newly isolated SSCs were purified and plated on MEF feeder (a), and were identified with IF staining using antibodies against PLZF (b), GFRA1 (c) and CDH1 (d). e Typical colonies of transformed cells from long-term cultured SSCs were exhibited. f-h Identification of transformed pluripotent cells using dual IF staining of OCT4 (f), SOX2 (g) and DAPI (h). i-j The morphology of ESCs (i) and ES-like cells derived from transformed pluripotent cells (j) on MEF feeder were exhibited. k-p The expression of pluripotent and germline markers (k. SSEA1, l. NANOG, m. merge; n. OCT4, o. MVH, p. merge) in ES-like cells were detected using IF staining. q The expression of germline and pluripotent markers were determined using RT-PCR (M. marker, 1. newly isolated SSCs, 2. GSPCs derived from long-term culture, 3. ES-like cells derived from GSPCs, 4. H2O). r-t The alkaline phosphatase activity was detected in ESCs (r), GSPCs (s) and ES-like cells (t). Scale bar = 20 μm