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

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

doi:10.1038/nbt.1580

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

Journal article

Peer reviewed

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

In-Hyun Park, George Q Daley, Sutheera Ratanasirintrawoot, Odelya Hartung, Yuin-Han Loh, Philip D Manos, Junsung Rho, Elayne M Chan, Justine D Miller, Hongguang Huo, …

Nature biotechnology, Vol.27(11), pp.1033-1037

2009-11

DOI: https://doi.org/10.1038/nbt.1580

PMID: 19826408

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.

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Collaboration types: Domestic collaboration
Citation topics: 1 Clinical & Life Sciences; 1.102 Stem Cell Research; 1.102.622 Pluripotency
Web Of Science research areas: Biotechnology & Applied Microbiology
ESI research areas: Biology & Biochemistry

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Title: Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells
Creators: In-Hyun Park - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School These authors contributed equally to this work
George Q Daley - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School Stem Cell Program, Children's Hospital Boston Harvard Stem Cell Institute Howard Hughes Medical Institute at Children's Hospital Boston Division of Hematology/Oncology, Brigham and Women's Hospital Manton Center for Orphan Disease Research, Children's Hospital Boston
Sutheera Ratanasirintrawoot - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston Biological and Biomedical Sciences, Harvard Medical School These authors contributed equally to this work
Odelya Hartung - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston
Yuin-Han Loh - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School
Philip D Manos - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston
Junsung Rho - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute
Elayne M Chan - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School These authors contributed equally to this work
Justine D Miller - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston
Hongguang Huo - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston
Tan A Ince - Department of Pathology, Brigham and Women's Hospital
Thorsten M Schlaeger - Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute Stem Cell Program, Children's Hospital Boston Harvard Stem Cell Institute
Publication Details: Nature biotechnology, Vol.27(11), pp.1033-1037
Language: English
Resource Type: Journal article
PMID: 19826408
Record Identifier: 991031614400402976