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Isolation of skin-derived precursors (SKPs) and differentiation and enrichment of their Schwann cell progeny

Nature Protocols volume 1, pages 2803–2812 (2006)Cite this article

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.

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Figure 1: SKPs grown in FGF2 and EGF.
Figure 2: Morphology of SKP-derived Schwann cells.

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Authors and Affiliations

  1. 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

  2. Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada

    Freda D Miller

  3. Department of Physiology, University of Toronto, Toronto, Ontario, Canada

    Freda D Miller

Authors
  1. Jeffrey A Biernaskie
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  2. Ian A McKenzie
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Correspondence to Freda D Miller.

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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.

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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

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