High-Yield Purification, Preservation, and Serial Transplantation of Human Satellite Cells

Steven M. Garcia; Stanley Tamaki; Solomon Lee; Alvin Wong; Anthony Jose; Joanna Dreux; Gayle Kouklis; Hani Sbitany; Rahul Seth; P. Daniel Knott; Chase Heaton; William R. Ryan; Esther A. Kim; Scott L. Hansen; William Y. Hoffman; Jason H. Pomerantz

Stem Cell Reports (Mar 2018)

High-Yield Purification, Preservation, and Serial Transplantation of Human Satellite Cells

Steven M. Garcia,
Stanley Tamaki,
Solomon Lee,
Alvin Wong,
Anthony Jose,
Joanna Dreux,
Gayle Kouklis,
Hani Sbitany,
Rahul Seth,
P. Daniel Knott,
Chase Heaton,
William R. Ryan,
Esther A. Kim,
Scott L. Hansen,
William Y. Hoffman,
Jason H. Pomerantz

Affiliations

Steven M. Garcia: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Stanley Tamaki: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Solomon Lee: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Alvin Wong: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Anthony Jose: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Joanna Dreux: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Gayle Kouklis: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Hani Sbitany: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Rahul Seth: Department of Otolaryngology - Head and Neck Surgery, University of California, San Francisco, CA 94143, USA
P. Daniel Knott: Department of Otolaryngology - Head and Neck Surgery, University of California, San Francisco, CA 94143, USA
Chase Heaton: Department of Otolaryngology - Head and Neck Surgery, University of California, San Francisco, CA 94143, USA
William R. Ryan: Department of Otolaryngology - Head and Neck Surgery, University of California, San Francisco, CA 94143, USA
Esther A. Kim: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Scott L. Hansen: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
William Y. Hoffman: Department of Surgery, Division of Plastic and Reconstructive Surgery, University of California, San Francisco, CA 94143, USA
Jason H. Pomerantz: Departments of Surgery and Orofacial Sciences, Division of Plastic and Reconstructive Surgery, Program in Craniofacial Biology, Eli and Edythe Broad Center of Regeneration Medicine, University of California, San Francisco, CA 94143, USA; Corresponding author

Journal volume & issue: Vol. 10, no. 3
pp. 1160 – 1174

Abstract

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Summary: Investigation of human muscle regeneration requires robust methods to purify and transplant muscle stem and progenitor cells that collectively constitute the human satellite cell (HuSC) pool. Existing approaches have yet to make HuSCs widely accessible for researchers, and as a result human muscle stem cell research has advanced slowly. Here, we describe a robust and predictable HuSC purification process that is effective for each human skeletal muscle tested and the development of storage protocols and transplantation models in dystrophin-deficient and wild-type recipients. Enzymatic digestion, magnetic column depletion, and 6-marker flow-cytometric purification enable separation of 104 highly enriched HuSCs per gram of muscle. Cryostorage of HuSCs preserves viability, phenotype, and transplantation potential. Development of enhanced and species-specific transplantation protocols enabled serial HuSC xenotransplantation and recovery. These protocols and models provide an accessible system for basic and translational investigation and clinical development of HuSCs. : Garcia and colleagues report methods for efficient purification of satellite cells from human skeletal muscle. They use their approaches to demonstrate stem cell functions of endogenous satellite cells and to make human satellite cells accessible for sharing among researchers. Keywords: human satellite cell purification, serial transplantation, satellite cell cryopreservation

Published in Stem Cell Reports

ISSN: 2213-6711 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Medicine (General); Science: Biology (General)
Website: http://www.cell.com/stem-cell-reports/home

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