Cytogenetic analysis confirmed that all 8 iPSC lines were karotypically normal. Gene expression of pluripotency markers and inactivation of Sendai-virus particles were confirmed in all iPSC lines by qRT-PCR as described by Park et al. [30] and Chan et al [31] (Panel A of S2 Fig). Protein expression of pluripotency markers was demonstrated by immunocytochemistry (Panel B of S2 Fig). To confirm the capacity of iPS cells to spontaneously differentiate into three germ layers, EBs were generated and maintained in suspension for spontaneous differentiation. Expression of genes specific for three germ layers was demonstrated by RT-PCR (Panel C of S2 Fig).
