Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells Export

@article{citeulike:6081479, abstract = {Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by enforced expression of transcription factors. Using serial live imaging of human fibroblasts undergoing reprogramming, we identified distinct colony types that morphologically resemble embryonic stem (ES) cells yet differ in molecular phenotype and differentiation potential. By analyzing expression of pluripotency markers, methylation at the OCT4 and NANOG promoters and differentiation into teratomas, we determined that only one colony type represents true iPS cells, whereas the others represent reprogramming intermediates. Proviral silencing and expression of TRA-1-60, DNMT3B and REX1 can be used to distinguish the fully reprogrammed state, whereas alkaline phosphatase, SSEA-4, GDF3, hTERT and NANOG are insufficient as markers. We also show that reprogramming using chemically defined medium favors formation of fully reprogrammed over partially reprogrammed colonies. Our data define molecular markers of the fully reprogrammed state and highlight the need for rigorous characterization and standardization of putative iPS cells.}, author = {Chan, Elayne M. and Ratanasirintrawoot, Sutheera and Park, In-Hyun and Manos, Philip D. and Loh, Yuin-Han and Huo, Hongguang and Miller, Justine D. and Hartung, Odelya and Rho, Junsung and Ince, Tan A. and Daley, George Q. and Schlaeger, Thorsten M.}, booktitle = {Nat Biotech}, citeulike-article-id = {6081479}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nbt.1580}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nbt.1580}, day = {11}, doi = {10.1038/nbt.1580}, issn = {1087-0156}, journal = {Nature Biotechnology}, keywords = {differentiation, stem\_cell}, month = {October}, number = {11}, pages = {1033--1037}, posted-at = {2009-11-06 20:00:40}, priority = {2}, publisher = {Nature Publishing Group}, title = {Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells}, url = {http://dx.doi.org/10.1038/nbt.1580}, volume = {27}, year = {2009} }

TY - JOUR ID - citeulike:6081479 L3 - citeulike-article-id:6081479 N2 - Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by enforced expression of transcription factors. Using serial live imaging of human fibroblasts undergoing reprogramming, we identified distinct colony types that morphologically resemble embryonic stem (ES) cells yet differ in molecular phenotype and differentiation potential. By analyzing expression of pluripotency markers, methylation at the OCT4 and NANOG promoters and differentiation into teratomas, we determined that only one colony type represents true iPS cells, whereas the others represent reprogramming intermediates. Proviral silencing and expression of TRA-1-60, DNMT3B and REX1 can be used to distinguish the fully reprogrammed state, whereas alkaline phosphatase, SSEA-4, GDF3, hTERT and NANOG are insufficient as markers. We also show that reprogramming using chemically defined medium favors formation of fully reprogrammed over partially reprogrammed colonies. Our data define molecular markers of the fully reprogrammed state and highlight the need for rigorous characterization and standardization of putative iPS cells. IS - 11 JF - Nature Biotechnology SN - 1087-0156 EP - 1037 TI - Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells PB - Nature Publishing Group VL - 27 T2 - Nat Biotech SP - 1033 KW - differentiation KW - stem_cell AU - Chan, Elayne AU - Ratanasirintrawoot, Sutheera AU - Park, In-Hyun AU - Manos, Philip AU - Loh, Yuin-Han AU - Huo, Hongguang AU - Miller, Justine AU - Hartung, Odelya AU - Rho, Junsung AU - Ince, Tan AU - Daley, George AU - Schlaeger, Thorsten PY - 2009/10/11/ UR - http://dx.doi.org/10.1038/nbt.1580 ER -