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Isolation and Culture of Porcine Neural Progenitor Cells from Embryos and Pluripotent Stem Cells

  • Mikkel A. Rasmussen
  • Vanessa J. Hall
  • Poul Hyttel
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1074)

Abstract

The isolation and culture of neural progenitor cells (NPCs) from pluripotent stem cells has facilitated in vitro mechanistic studies of diseases related to the nervous system, as well as discovery of new medicine. In addition, NPCs are envisioned to play a crucial role in future cell replacement therapy. The pig has become recognized as an important large animal model and establishment of in vitro-derived porcine NPCs would allow for preclinical safety testing by transplantation in a porcine biomedical model. In this chapter, a detailed method for isolation and in vitro culture of porcine NPCs from porcine embryos or induced pluripotent stem cells is presented. The neural induction is performed in coculture and the isolation of rosette structures is carried out manually to ensure a homogenous population of NPCs. Using this method, multipotent NPCs can be obtained in approximately 1 month. The cells have the potential of long-term culture and the ability to differentiate into neural and glial cells.

Key words

Neural progenitor cells Embryos Induced pluripotent stem cells Neurons and glia Transplantation 

Notes

Acknowledgement

The Danish National Advanced Technology Foundation (HTF) 047-2011-1, The Danish Council for Independent Research, Natural Sciences (FNU), 11-106627, and The Danish Council for Strategic Research (DSF)—The Program Commission on Individuals, Disease and Society.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Mikkel A. Rasmussen
    • 1
  • Vanessa J. Hall
    • 1
  • Poul Hyttel
    • 1
  1. 1.Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical SciencesUniversity of CopenhagenFrederiksberg CDenmark

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