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Isolation of Myogenic Stem Cells from Cultures of Cryopreserved Human Skeletal Muscle

Zheng, B and Chen, CW and Li, G and Thompson, SD and Poddar, M and Péault, B and Huard, J (2012) Isolation of Myogenic Stem Cells from Cultures of Cryopreserved Human Skeletal Muscle. Cell Transplantation, 21 (6). 1087 - 1093. ISSN 0963-6897

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Abstract

We demonstrate that subpopulations of adult human skeletal muscle-derived stem cells, myogenic endothelial cells (MECs), and perivascular stem cells (PSCs) can be simultaneously purified by fluorescence-activated cell sorting (FACS) from cryopreserved human primary skeletal muscle cell cultures (cryo-hPSMCs). For FACS isolation, we utilized a combination of cell lineage markers: the myogenic cell marker CD56, the endothelial cell marker UEA-1 receptor (UEA-1R), and the perivascular cell marker CD146. MECs expressing all three cell lineage markers (CD56+UEA-1R+CD146+/CD45+) and PSCs expressing only CD146 (CD146+/CD45+CD56+UEA-1R+) were isolated by FACS. To evaluate their myogenic capacities, the sorted cells, with and without expansion in culture, were transplanted into the cardiotoxin-injured skeletal muscles of immunodeficient mice. The purified MECs exhibited the highest regenerative capacity in the injured mouse muscles among all cell fractions tested, while PSCs remained superior to myoblasts and the unpurified primary skeletal muscle cells. Our findings show that both MECs and PSCs retain their high myogenic potentials after in vitro expansion, cryopreservation, and FACS sorting. The current study demonstrates that myogenic stem cells are prospectively isolatable from long-term cryopreserved primary skeletal muscle cell cultures. We emphasize the potential application of this new approach to extract therapeutic stem cells from human muscle cells cryogenically banked for clinical purposes. © 2012 Cognizant Comm. Corp.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Zheng, B
Chen, CW
Li, G
Thompson, SD
Poddar, Mmip45@pitt.eduMIP45
Péault, B
Huard, J
Centers: Other Centers, Institutes, or Units > Stem Cell Research Center
Date: 3 September 2012
Date Type: Publication
Journal or Publication Title: Cell Transplantation
Volume: 21
Number: 6
Page Range: 1087 - 1093
DOI or Unique Handle: 10.3727/096368912x636876
Schools and Programs: School of Medicine > Orthopaedic Surgery
School of Medicine > Pediatrics
Swanson School of Engineering > Bioengineering
Refereed: Yes
ISSN: 0963-6897
MeSH Headings: Adolescent; Adult; Aged; Animals; Antigens, CD146--metabolism; Antigens, CD56--metabolism; Cell Differentiation; Cell Lineage; Cells, Cultured; Child; Child, Preschool; Cryopreservation; Endothelial Cells--cytology; Female; Flow Cytometry; Humans; Male; Mice; Mice, SCID; Middle Aged; Muscle, Skeletal--cytology; Regeneration--physiology; Stem Cell Transplantation; Stem Cells--cytology; Stem Cells--metabolism; Transcription Factors--metabolism; Ubiquitin-Conjugating Enzymes--metabolism; Young Adult
PubMed ID: 22472558
Date Deposited: 04 Apr 2014 15:36
Last Modified: 29 Jan 2019 15:55
URI: http://d-scholarship.pitt.edu/id/eprint/20848

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