Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells英文文献.pdfVIP

l e t t e r s live cell imaging distinguishes bona fide human iPs cells from partially reprogrammed cells Elayne M Chan1,2,10, Sutheera Ratanasirintrawoot1,3,4,10, In-Hyun Park1,2,10, Philip D Manos1,3, Yuin-Han Loh1,2, 1,3 1,3 1,3 1 5 1–3,6–9 Hongguang Huo , Justine D Miller , Odelya Hartung , Junsung Rho , Tan A Ince , George Q Daley Thorsten M Schlaeger1,3,6 Somatic cells can be reprogrammed into induced pluripotent destined to become bona fide human iPS cell lines downregulate stem (iPS) cells by enforced expression of transcription factors. the fibroblast marker CD13, upregulate the pluripotency markers Using serial live imaging of human fibroblasts undergoing SSEA-4 and TRA-1-60, silence the transgenes, reactivate expres- reprogramming, we identified distinct colony types that sion of endogenous NANOG and transiently assume a ‘Hoechstdim’ morphologically resemble embryonic stem (ES) cells yet phenotype. Our results suggest that analysis of multiple markers is differ in molecular phenotype and differentiation potential. essential to distinguish the fully reprogrammed state from partially By analyzing expression of pluripotency markers, methylation reprogrammed intermediates. at the OCT4 and NANOG promoters and differentiation into We first studied reprogramming of human ES cell–derived fibro- teratomas, we determined that only one colony type represents blasts, which, like mouse embryonic fibroblasts (MEFs),