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. Author manuscript; available in PMC: 2015 Jan 28.
Published in final edited form as: Stem Cells. 2013 May;31(5):895–905. doi: 10.1002/stem.1323

Figure 1. GMPs are a suitable source cell type for imaging pluripotency induction.

Figure 1

GMPs from adult bone marrow of mice harboring Rosa26-rtTA and Oct4-GFP were transduced with a polycistronic lentivirus expressing Oct-4, Sox2, Klf4 and c-Myc (OSKM) and plated on inactivated feeder cells in mESC culture conditions supplemented with of 2 μg/ml dox. Representative images of Oct-4 GFP+ colonies after 3, 5 and 8 days are shown. A) Cells that were not reprogrammed (red arrows) show typical non-adherent, round morphology. B) By day 5, large, tightly compact colonies were present. C) Without passaging, spontaneous differentiation occurs in some colonies by day 8, as indicated by the loss of Oct-4 GFP expression at the periphery of the colony (red arrows). D) Dox-independent Oct-4 GFP+ single colonies were picked and injected into B6 albino blastocysts. High degree chimeras were observed as indicated by their mosaic coat color. Upon breeding with albino females, the chimeric males fathered completely agouti pups indicating germline transmission. E) A representative low power view of a reprogramming culture 5 days after dox addition. Note the lack of partially GFP+ colonies.