Figure 2.

Characterization of CM- and CF-derived iPS colonies. (a) Number of iPS colonies/35 mm-dish formed on MEFs after 15 days from transduction of CMs and CFs with OSK reprogramming factors. ^*P<0.005: t-test (N=5 experiments for each clone). (b) Proliferation of post-natal CMs before and after transduction with OSK factors (left panel). BrdU labeling in a colony 15 days after sorting (right panel). Scale bar represents 200 μm. ^*P<0.005: t-test (N=5 experiments for each clone). (c) Morphology of CM- and CF-derived cells after 15 days from reprogramming (upper and lower left panels). Morphology of CM- and CF-derived iPS colonies at passages 2 and 20 (upper and lower right panels). Scale bars represent 200 μm. (d) Example of a 12-day-old CM-derived iPS clone stained for SSEA-1 (upper left panel); ALP expression in neonatal CM-derived iPS cells 15 days after transduction with OSK vectors (upper right panel); Oct4 and DAPI staining (lower right and left panels, respectively) in a colony formed from CMs. Scale bars represent 200 μm. (e) Teratoma formation 4 weeks after injection of CM-derived iPS cells in NOD-scid mice. Hematoxylin/eosin-stained sections demonstrating the presence of all three germ layers. (f) Normal karyotype of neonatal CM-derived iPS cells. (g) Two-week-old chimeric mouse, derived from a C57BL/6 blastocyst injected with CM-derived iPS cells obtained from a white CD-1 mouse, between two C57BL/6 wild-type littermates. CM-derived iPS cells are responsible for the agouti coat color. (h) qRT-PCR analysis of embryonic- and cardiac-marker genes in mES cells and CMs transduced with OSK factors (N=5 experiments for each clone)