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. Author manuscript; available in PMC: 2017 Mar 3.
Published in final edited form as: Cell Stem Cell. 2016 Feb 11;18(3):354–367. doi: 10.1016/j.stem.2015.12.001

Figure 1. Screening for iCPC Inducing Factors and Optimal Culture Conditions.

Figure 1

(A) Schematic representation of experimental design depicting direct reprogramming of adult fibroblasts to iCPCs by defined factors and culture conditions, expansion of iCPCs, and in vitro as well as in vivo differentiation of iCPCs into cardiac-lineage cells. (B) Infection with a combination of 11 cardiac factors induced Nkx2.5-EYFP expression in AC Fibs only after dox induction. (C) Strategy to test the impact of culture conditions on F11 reprogramming efficiency as well as the ability of EYFP+ cells to maintain a proliferative state. (D) Number of EYFP+ colonies formed (per 50,000 starting cells) in the respective culture conditions (**p<0.01, *p<0.05). (E) Impact of culture conditions on EYFP+ colonies expanded up to 5 psgs scoring for EYFP+ expression and proliferative ability (Dox only: n=8, Dox+LIF: n=3, Dox+BIO: n=4, Dox+LIF+BIO: n=9). (L=LIF, B=BIO). (F) F11 iCPCs maintained EYFP expression and proliferative ability for at least 30 psgs after dox withdrawal. (G) Population doubling time for psg 10 (P10) and psg 20 (P20) F11 iCPCs as compared to uninfected AC Fibs (n=3). Data presented as mean. Error bars = SEM. Scale bar = 100 μm in B, 200 μm in F. See also Figure S1, S2 and Movie S1.