Expression of pluripotency markers by iPSCs prepared with LIF or bFGF.

(A) Immunofluorescence staining of iPSCs prepared with LIF or bFGF on day 8. Antibodies against NANOG and TRA1-60 were used, and DNA was counterstained with Hoechst 33342. Bar, 100 µm in (A) and (C). (B) Formation of iPSC colonies that were double-positive for NANOG and TRA1-60 with LIF, bFGF, or no cytokines. Cells (1.7×10⁴) were seeded in each well, and iPSC colonies were counted on day 10. Mean + standard deviation (SD) was obtained from three independent experiments. (C) Immunofluorescence staining of iPSCs prepared with LIF or bFGF on day 8. Antibodies against SSEA4 and NANOG were used. (D) Formation of iPSC colonies that were double-positive for SSEA4 and NANOG with bFGF, LIF, or no cytokines. Cells were seeded and iPSC colonies counted as described in (B).