Magnetic-activated cell sorting for the fast and efficient separation of human and rodent Schwann cells from mixed cell populations

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

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages87-109
Number of pages23
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1739
ISSN (Print)1064-3745

Keywords

  • Cell sorting
  • Fibroblasts
  • Magnetic bead separation
  • O1
  • O4
  • p75
  • Peripheral nerve
  • Primary cultures
  • Schwann cells
  • Thy-1

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    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 Methods in Molecular Biology (pp. 87-109). (Methods in Molecular Biology; Vol. 1739). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7649-2_6