A drug-inducible system for direct reprogramming of human somatic cells to pluripotency.
Cell Stem Cell
; 3(3): 346-353, 2008 Sep 11.
Article
em En
| MEDLINE
| ID: mdl-18786421
ABSTRACT
Current approaches to reprogram human somatic cells to pluripotent iPSCs utilize viral transduction of different combinations of transcription factors. These protocols are highly inefficient because only a small fraction of cells carry the appropriate number and stoichiometry of proviral insertions to initiate the reprogramming process. Here we have generated genetically homogeneous "secondary" somatic cells, which carry the reprogramming factors as defined doxycycline (DOX)-inducible transgenes. These cells were obtained by infecting fibroblasts with DOX-inducible lentiviruses, isolating "primary" iPSCs in the presence of the drug, and finally differentiating to "secondary" fibroblasts. When "secondary" fibroblast lines were cultured in the presence of DOX without further viral infection, up to 2% of the cells were reprogrammed to pluripotent "secondary" human iPSCs. This system will facilitate the characterization of the reprogramming process and provides a unique platform for genetic or chemical screens to enhance reprogramming or replace individual factors.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Técnicas Citológicas
/
Células-Tronco Pluripotentes
/
Reprogramação Celular
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Desdiferenciação Celular
Limite:
Humans
Idioma:
En
Revista:
Cell Stem Cell
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos