Transdifferentiation of Mouse Fibroblasts and Hepatocytes to Functional Neurons

Part of the Methods in Molecular Biology book series (MIMB, volume 1150)


Nuclear reprogramming by defined transcription factors became of broad interest in 2006 with the work of Takahashi and Yamanaka (Cell 126:663–676, 2006), but the first example of cell fate reshaping via ectopic expression of transcription factor was provided back in 1987 when Davis and colleagues induced features of a muscle cell in fibroblast using the muscle transcription factor MyoD (Davis et al., Cell 51:987–1000, 1987). In 2010 our laboratory described how forced expression of the three neuronal transcription factors Ascl1, Brn2, and Myt1l rapidly converts mouse fibroblasts into neuronal cells that exhibit biochemical and electrophysiological properties of neurons. We named these cells induced neuronal cells (iN cells) (Vierbuchen et al., Nature 463:1035–1041, 2010; Vierbuchen and Wernig, Nat Biotechnol 29:892–907, 2011). Interestingly, iN cells can also be derived from defined endodermal cells such as primary hepatocytes, suggesting the existence of a more general reprogramming paradigm (Marro et al., Cell Stem Cell 9:374–382, 2011). In this chapter we describe the detailed methods used to attain the direct conversion.

Key words

Nuclear reprogramming Fibroblast Hepatocyte Induced neuronal cells 


  1. 1.
    Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663–676PubMedCrossRefGoogle Scholar
  2. 2.
    Davis RL, Weintraub H, Lassar AB (1987) Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51:987–1000PubMedCrossRefGoogle Scholar
  3. 3.
    Vierbuchen T et al (2010) Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463:1035–1041PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Vierbuchen T, Wernig M (2011) Direct lineage conversions: unnatural but useful? Nat Biotechnol 29:892–907PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Marro S et al (2011) Direct lineage conversion of terminally differentiated hepatocytes to functional neurons. Cell Stem Cell 9:374–382PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Gehring W (1967) Clonal analysis of determination dynamics in cultures of imaginal disks in Drosophila melanogaster. Dev Biol 16:438–456PubMedCrossRefGoogle Scholar
  7. 7.
    Hadorn E (1966) Constancy, variation and type of determination and differentiation in cells from male genitalia rudiments of Drosophila melanogaster in permanent culture in vivo. Dev Biol 13:424–509PubMedCrossRefGoogle Scholar
  8. 8.
    Pagan R et al (1995) Epithelial-mesenchymal transition in cultured neonatal hepatocytes. Hepatology 21:820–831PubMedCrossRefGoogle Scholar
  9. 9.
    Gossen M et al (1995) Transcriptional activation by tetracyclines in mammalian cells. Science 268:1766–1769PubMedCrossRefGoogle Scholar
  10. 10.
    Tiscornia G, Singer O, Verma IM (2006) Production and purification of lentiviral vectors. Nat Protoc 1:241–245PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Pathology, Institute for Stem Cell Biology and Regenerative MedicineStanford University School of MedicineStanfordUSA

Personalised recommendations