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Magnetic-Activated Cell Sorting for the Fast and Efficient Separation of Human and Rodent Schwann Cells from Mixed Cell Populations

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Schwann Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1739))

Abstract

To date, magnetic-activated cell sorting (MACS) remains a powerful method to isolate distinct cell populations based on differential cell surface labeling. Optimized direct and indirect MACS protocols for cell immunolabeling are presented here as methods to divest Schwann cell (SC) cultures of contaminating cells (specifically, fibroblast cells) and isolate SC populations at different stages of differentiation. This chapter describes (1) the preparation of single-cell suspensions from established human and rat SC cultures, (2) the design and application of cell selection strategies using SC-specific (p75NGFR, O4, and O1) and fibroblast-specific (Thy-1) markers, and (3) the characterization of both the pre- and post-sorting cell populations. A simple protocol for the growth of hybridoma cell cultures as a source of monoclonal antibodies for cell surface immunolabeling of SCs and fibroblasts is provided as a cost-effective alternative for commercially available products. These steps allow for the timely and efficient recovery of purified SC populations without compromising the viability and biological activity of the cells.

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Acknowledgments

We are grateful for the technical assistance provided by Dr. Ketty Bacallao and Ms. Blanche Kuo. We thank Dr. James Guest for critically reviewing the manuscript. The work presented in this chapter was generously supported by the NIH-NINDS (NS084326), The Craig Neilsen Foundation (339576), The Miami Project to Cure Paralysis, and The Buoniconti Fund. The authors declare no conflicts of interest with the contents of this article.

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Author notes

  1. Kristine M. Ravelo, Natalia D. Andersen contributed equally to this work.

Authors and Affiliations

  1. The Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, USA

    Kristine M. Ravelo, Natalia D. Andersen & Paula V. Monje

Authors
  1. Kristine M. Ravelo
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  2. Natalia D. Andersen
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  3. Paula V. Monje
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Corresponding author

Correspondence to Paula V. Monje .

Editor information

Editors and Affiliations

  1. Department of Neurological Surgery, University of Miami, Miami, Florida, USA

    Paula V. Monje

  2. Department of Biological Sciences, Rutgers University, Newark, New Jersey, USA

    Haesun A. Kim

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Ravelo, K.M., Andersen, N.D., Monje, P.V. (2018). Magnetic-Activated Cell Sorting for the Fast and Efficient Separation of Human and Rodent Schwann Cells from Mixed Cell Populations. In: Monje, P., Kim, H. (eds) Schwann Cells. Methods in Molecular Biology, vol 1739. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7649-2_6

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  • DOI: https://doi.org/10.1007/978-1-4939-7649-2_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7648-5

  • Online ISBN: 978-1-4939-7649-2

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