Format

Send to

Choose Destination
See comment in PubMed Commons below
Nat Protoc. 2014 Apr;9(4):871-81. doi: 10.1038/nprot.2014.056. Epub 2014 Mar 20.

Direct conversion of mouse fibroblasts into induced neural stem cells.

Author information

  • 11] Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul, Republic of Korea. [2] Department of Animal Biotechnology, Konkuk University, Seoul, Republic of Korea. [3].
  • 21] Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany. [2].
  • 31] Division for Neurodegenerative Diseases, Department of Neurology, Dresden University of Technology, Dresden, Germany. [2] Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany.
  • 41] Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany. [2] Group of Computational Biology and Bioinformatics, Biodonostia Health Research Institute, San Sebastián, Spain. [3] IKERBASQUE, Basque Foundation for Science, Bilbao, Spain.
  • 5Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul, Republic of Korea.
  • 6Department of Animal Biotechnology, Konkuk University, Seoul, Republic of Korea.
  • 7Department of Internal Medicine, School of Medicine, Konkuk University, Seoul, Republic of Korea.
  • 8Department of Microbiology, School of Medicine, Konkuk University, Seoul, Republic of Korea.
  • 91] Division for Neurodegenerative Diseases, Department of Neurology, Dresden University of Technology, Dresden, Germany. [2] Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany. [3] Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany.
  • 101] Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany. [2] Medical Faculty, University of Münster, Münster, Germany.
  • 11Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany.

Abstract

Terminally differentiated cells can be directly converted into different types of somatic cells by using defined factors, thus circumventing the pluripotent state. However, low reprogramming efficiency, along with the absence of proliferation of some somatic cell types, makes it difficult to generate large numbers of cells with this method. Here we describe a protocol to directly convert mouse fibroblasts into self-renewing induced neural stem cells (iNSCs) that can be expanded in vitro, thereby overcoming the limitations associated with low reprogramming efficiency. The four transcription factors required for direct conversion into iNSCs (Sox2, Klf4, Myc (also known as c-Myc) and Pou3f4 (also known as Brn4)) do not generate a pluripotent cell state, and thus the risk for tumor formation after transplantation is reduced. By following the current protocol, iNSCs are observed 4-5 weeks after transduction. Two additional months are required to establish clonal iNSC cell lines that exhibit retroviral transgene silencing and that differentiate into neurons, astrocytes and oligodendrocytes.

PMID:
24651499
DOI:
10.1038/nprot.2014.056
[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Nature Publishing Group
    Loading ...
    Support Center