A microRNA-based system for selecting and maintaining the pluripotent state in human induced pluripotent stem cells

Bruno Di Stefano, Sara Martina Maffioletti, Bernhard Gentner, Federica Ungaro, Giulia Schira, Luigi Naldini, Vania Broccoli

Research output: Contribution to journalArticle

Abstract

Induced pluripotent stem cell (iPSC) technology has provided researchers with a unique tool to derive disease-specific stem cells for the study and possible treatment of degenerative disorders with autologous cells. The low efficiency and heterogeneous nature of reprogramming is a major impediment to the generation of personalized iPSC lines. Here, we report the generation of a lentiviral system based on a microRNA-regulated transgene that enables for the efficient selection of mouse and human pluripotent cells. This system relies on the differential expression pattern of the mature form of microRNA let7a in pluripotent versus committed or differentiated cells. We generated microRNA responsive green fluorescent protein and Neo reporters for specific labeling and active selection of the pluripotent cells in any culture condition. We used this system to establish Rett syndrome and Parkinson's disease human iPSCs. The presented selection procedure represents a straightforward and powerful tool for facilitating the derivation of patient-specific iPSCs.

Original languageEnglish
Pages (from-to)1684-1695
Number of pages12
JournalStem Cells
Volume29
Issue number11
DOIs
Publication statusPublished - Nov 2011

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Keywords

  • Genetic reprogramming
  • Induced pluripotent stem cells
  • Let7a
  • Lin28
  • MicroRNA
  • Parkinson's disease
  • Rett syndrome

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

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