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ABSTRACT This protocol describes methods of isolating skin-derived precursors (SKPs) from rodent and human skin, and for generating and enriching Schwann cells from rodent SKPs. SKPs are

isolated as a population of non-adherent cells from the dermis that proliferate and self-renew as floating spheres in response to fibroblast growth factor 2 (FGF2) and epidermal growth

factor (EGF). Their differentiation into Schwann cells and subsequent enrichment of these differentiated progeny involves culturing SKPs as adherent cells in the absence of FGF2 and EGF, but

in the presence of neuregulins, and then mechanically isolating the Schwann cell colonies using cloning cylinders. Methods for expanding and characterizing these Schwann cells are provided.

Generation of primary SKPs takes approximately 2 weeks, while differentiation of Schwann cells requires an additional 4–6 weeks. Access through your institution Buy or subscribe This is a

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Google Scholar  Download references AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Developmental Biology Group, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

Jeffrey A Biernaskie, Ian A McKenzie, Jean G Toma & Freda D Miller * Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada Freda D Miller *

Department of Physiology, University of Toronto, Toronto, Ontario, Canada Freda D Miller Authors * Jeffrey A Biernaskie View author publications You can also search for this author inPubMed 

Google Scholar * Ian A McKenzie View author publications You can also search for this author inPubMed Google Scholar * Jean G Toma View author publications You can also search for this

author inPubMed Google Scholar * Freda D Miller View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Freda D Miller.

ETHICS DECLARATIONS COMPETING INTERESTS Dr. Freda Miller is a consultant for Aggregate Therapeutics, a biotechnology company who has licensed patents covering SKPs technology from McGill

University, who owns the technology. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Biernaskie, J., McKenzie, I., Toma, J. _et al._ Isolation of

skin-derived precursors (SKPs) and differentiation and enrichment of their Schwann cell progeny. _Nat Protoc_ 1, 2803–2812 (2006). https://doi.org/10.1038/nprot.2006.422 Download citation *

Published: 25 January 2007 * Issue Date: December 2006 * DOI: https://doi.org/10.1038/nprot.2006.422 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this

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