Unit

UNIT 2D.3 Long-Term Multilayer Adherent Network (MAN) Expansion, Maintenance, and Characterization, Chemical and Genetic Manipulation, and Transplantation of Human Fetal Forebrain Neural Stem Cells

  1. Dustin R. Wakeman1,2,
  2. Martin R. Hofmann2,
  3. D. Eugene Redmond Jr.3,
  4. Yang D. Teng4,5,
  5. Evan Y. Snyder1,2

Published Online: 1 MAY 2009

DOI: 10.1002/9780470151808.sc02d03s9

Current Protocols in Stem Cell Biology

Current Protocols in Stem Cell Biology

How to Cite

Wakeman, D. R., Hofmann, M. R., Redmond, D. E., Teng, Y. D. and Snyder, E. Y. 2009. Long-Term Multilayer Adherent Network (MAN) Expansion, Maintenance, and Characterization, Chemical and Genetic Manipulation, and Transplantation of Human Fetal Forebrain Neural Stem Cells. Current Protocols in Stem Cell Biology. 9:D:2D.3:2D.3.1–2D.3.77.

Author Information

  1. 1

    University of California at San Diego, La Jolla, California

  2. 2

    The Burnham Institute for Medical Research, La Jolla, California

  3. 3

    Yale University School of Medicine, New Haven, Connecticut

  4. 4

    Harvard Medical School, Brigham & Women's Hospital and Spaulding Rehabilitation Hospital, Boston, Massachusetts

  5. 5

    Veterans Affairs Boston Healthcare System, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2009
  2. Published Print: SEP 2009

Abstract

Human neural stem/precursor cells (hNSC/hNPC) have been targeted for application in a variety of research models and as prospective candidates for cell-based therapeutic modalities in central nervous system (CNS) disorders. To this end, the successful derivation, expansion, and sustained maintenance of undifferentiated hNSC/hNPC in vitro, as artificial expandable neurogenic micro-niches, promises a diversity of applications as well as future potential for a variety of experimental paradigms modeling early human neurogenesis, neuronal migration, and neurogenetic disorders, and could also serve as a platform for small-molecule drug screening in the CNS. Furthermore, hNPC transplants provide an alternative substrate for cellular regeneration and restoration of damaged tissue in neurodegenerative disorders such as Parkinson's disease and Alzheimer's disease. Human somatic neural stem/progenitor cells (NSC/NPC) have been derived from a variety of cadaveric sources and proven engraftable in a cytoarchitecturally appropriate manner into the developing and adult rodent and monkey brain while maintaining both functional and migratory capabilities in pathological models of disease. In the following unit, we describe a new procedure that we have successfully employed to maintain operationally defined human somatic NSC/NPC from developing fetal, pre-term post-natal, and adult cadaveric forebrain. Specifically, we outline the detailed methodology for in vitro expansion, long-term maintenance, manipulation, and transplantation of these multipotent precursors. Curr. Protoc. Stem Cell Biol. 9:2D.3.1-2D.3.77. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • human neural stem/progenitor cell;
  • NPC;
  • NSC;
  • culture;
  • fetal/adult forebrain;
  • subventricular zone;
  • neurogenesis;
  • niche;
  • multilayer adherent network;
  • MAN assay;
  • protocols;
  • manipulation techniques;
  • characterization;
  • in vitro;
  • derivation;
  • expansion;
  • maintenance;
  • SPIO;
  • Feridex;
  • lentivirus;
  • BrdU;
  • labeling