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

Elayne M. Chan, Sutheera Ratanasirintrawoot, In Hyun Park, Philip D. Manos, Yuin Han Loh, Hongguang Huo, Justine D. Miller, Odelya Hartung, Junsung Rho, Tan A. Ince, George Q. Daley, Thorsten M. Schlaeger

Research output: Contribution to journalArticlepeer-review

383 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)1033-1037
Number of pages5
JournalNature Biotechnology
Issue number11
StatePublished - Nov 2009
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering


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