Methods ARTICLE

Front. Neurosci., 14 July 2011 | doi: 10.3389/fnins.2011.00089

A protocol for isolation and enriched monolayer cultivation of neural precursor cells from mouse dentate gyrus

  • 1 Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA
  • 2 CRTD, Center for Regenerative Therapies Dresden, DFG-Research Center for Regenerative Therapies Dresden, Dresden, Germany
  • 3 DZNE, German Center for Neurodegenerative Diseases, Dresden, Germany
  • 4 Department of Neurology, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany

In vitro assays are valuable tools to study the characteristics of adult neural precursor cells under controlled conditions with a defined set of parameters. We here present a detailed protocol based on our previous original publication (Babu et al., 2007) to isolate neural precursor cells from the hippocampus of adult mice and maintain and propagate them as adherent monolayer cultures. The strategy is based on the use of Percoll density gradient centrifugation to enrich precursor cells from the micro-dissected dentate gyrus. Based on the expression of Nestin and Sox2, a culture-purity of more than 98% can be achieved. The cultures are expanded under serum-free conditions in Neurobasal A medium with addition of the mitogens Epidermal growth factor and Fibroblast growth factor 2 as well as the supplements Glutamax-1 and B27. Under differentiation conditions, the precursor cells reliably generate approximately 30% neurons with appropriate morphological, molecular, and electrophysiological characteristics that might reflect granule cell properties as their in vivo counterpart. We also highlight potential modifications to the protocol.

Keywords: hippocampus, progenitor cell, precursor cell, adult neurogenesis, in vitro

Citation: Babu H, Claasen J-H, Kannan S, Rünker AE, Palmer T and Kempermann G (2011) A protocol for isolation and enriched monolayer cultivation of neural precursor cells from mouse dentate gyrus. Front. Neurosci. 5:89. doi: 10.3389/fnins.2011.00089

Received: 11 March 2011; Accepted: 28 June 2011;
Published online: 14 July 2011.

Edited by:

Silvia De Marchis, University of Turin, Italy

Reviewed by:

Silvia De Marchis, University of Turin, Italy
D. Chichung Lie, Helmholtz Zentrum Munchen German Research Center for Environmental Health, Germany

Copyright: © 2011 Babu, Claasen, Kannan, Rünker, Palmer and Kempermann. This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and other Frontiers conditions are complied with.

*Correspondence: Gerd Kempermann, DFG Research Center for Regenerative Therapies Dresden, Center for Regenerative Therapies Dresden, Tatzberg 47–49, 01307 Dresden, Germany. e-mail: gerd.kempermann@crt-dresden.de

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