Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach

Kaiwen Peng, David Sant, Natalia Andersen, Risset Silvera, Vladimir Camarena, Gonzalo Piñero, Regina Graham, Aisha Khan, Xiao Ming Xu, Gaofeng Wang, Paula V. Monje

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17 Scopus citations


Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.

Original languageEnglish
Article number18433
JournalScientific Reports
Issue number1
StatePublished - Dec 1 2020

Bibliographical note

Funding Information:
We thank Ketty Bacallao, Kristine Ravelo and Blanche Kuo for assisting with cell culture and RNA isolation, Yan Shi for automated fluorescence microscopy, and Maxwell Donaldson for preparation of cell stocks. We greatly acknowledge the contribution of Patrick Wood and Mary Bunge for facilitating access to primary cells and cell lines, and Tamara Weiss for providing critical review of the manuscript. Illustrations were prepared by Christopher Brown and Patti Raley contributed with English editing. This work was supported by The Craig H. Neilsen Foundation (grant 339576 to P.V.M. and G.W), The Miami Project to Cure Paralysis and the Buoniconti Fund (to P.V.M), and the Indiana Traumatic Spinal Cord and Brain Injury Research Fund from the Indiana State Department of Health (to P.V.M and X.M.X). X.M.X received support from NIH 1R01 100531, 1R01 NS103481, and Merit Review Award I01 BX002356, I01 BX003705, I01 RX002687 from the U.S. Department of Veterans Affairs. The authors declare no conflicts of interest with the contents of this article.

Publisher Copyright:
© 2020, The Author(s).

ASJC Scopus subject areas

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