A Drug-inducible System for Direct Reprogramming of Human Somatic Cells to Pluripotency

Overview

Journal Cell Stem Cell

Publisher Cell Press

Specialty Cell Biology

Date 2008 Sep 13

PMID 18786421

Citations 187

Authors

Dirk Hockemeyer

Frank Soldner

Elizabeth G Cook

Qing Gao

Maisam Mitalipova

Rudolf Jaenisch

Affiliations

Soon will be listed here.

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.

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