Reprogramming to pluripotency through a somatic stem cell intermediate

PLoS One. 2013 Dec 27;8(12):e85138. doi: 10.1371/journal.pone.0085138. eCollection 2013.

Abstract

Transcription factor-based reprogramming can lead to the successful switching of cell fates. We have recently reported that mouse embryonic fibroblasts (MEFs) can be directly reprogrammed into induced neural stem cells (iNSCs) after the forced expression of Brn4, Sox2, Klf4, and Myc. Here, we tested whether iNSCs could be further reprogrammed into induced pluripotent stem cells (iPSCs). The two factors Oct4 and Klf4 were sufficient to induce pluripotency in iNSCs. Immunocytochemistry and gene expression analysis showed that iNSC-derived iPSCs (iNdiPSCs) are similar to embryonic stem cells at the molecular level. In addition, iNdiPSCs could differentiate into cells of all three germ layers, both in vitro and in vivo, proving that iNdiPSCs are bona fide pluripotent cells. Furthermore, analysis of the global gene expression profile showed that iNdiPSCs, in contrast to iNSCs, do not retain any MEF transcriptional memory even at early passages after reprogramming. Overall, our results demonstrate that iNSCs can be reprogrammed to pluripotency and suggest that cell fate can be redirected numerous times. Importantly, our findings indicate that the induced pluripotent cell state may erase the donor-cell type epigenetic memory more efficiently than other induced somatic cell fates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation*
  • Cellular Reprogramming*
  • Epigenesis, Genetic / genetics
  • Gene Expression Regulation*
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism*
  • Mice
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism*
  • Transcription Factors / biosynthesis*
  • Transcription Factors / genetics

Substances

  • Transcription Factors

Associated data

  • GEO/GSE44284

Grant support

A.G.M. and U.T. are supported by a fellowship of the International Max Planck Research School, Molecular Biomedicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.