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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 5. Cardiac Factors Stably Reprogram Adult Lung and Adult Tail-tip Fibroblasts into Proliferative and Multipotent iCPCs.

Figure 5

(A) Number of Nkx2.5-EYFP+ colonies produced (per 50,000 seeded cells) after infection of adult lung & adult tail-tip fibroblasts with 11 or 5 factors and culture in iCPC induction medium for 3 weeks (n=4). (L=LIF, B=BIO). (B) EYFP+ cells reprogrammed using 5 factors could be stably expanded without doxycycline for at least 10 psgs (Lung) and 5 psgs (Tail). (C) Immunolabeling revealed F5 lung-iCPCs & tail-tip-iCPCs had nuclear localization of CPC TFs (merged images are depicted, Red = Nkx2.5, Green = Irx4, Blue = DAPI), quantified in (D). (E&F) Flow cytometry analysis revealed that F5 lung-iCPCs & tail-tip-iCPCs expressed cell surface markers associated with CPCs (n=3). (G) F5 lung-iCPCs showed a normal diploid karyotype. (H) Lung-iCPCs were multipotent and differentiated into CMs (cardiac actin, α-actinin, MLC-2v, α/β MHC), SMs (SM-MHC) and ECs (CD31). Note highly organized sarcomere staining for α-actinin. (I) Tail-tip-iCPCs were multipotent and differentiated into cardiomyocytes (cardiac actin, α-actinin, α/β MHC), smooth muscle cells (SM-MHC) and endothelial cells (CD31). Note organized sarcomere staining for α-actinin. Data presented as mean, error bars = SEM. Scale bars = 100μm. See also Figure S2, S3.