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Isolation of Muscle-Derived Stem/Progenitor Cells Based on Adhesion Characteristics to Collagen-Coated Surfaces

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Stem Cells and Aging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 976))

Abstract

Our lab developed and optimized a method, known as the modified pre-plate technique, to isolate stem/progenitor cells from skeletal muscle. This method separates different populations of myogenic cells based on their propensity to adhere to a collagen I-coated surface. Based on their surface markers and stem-like properties, including self-renewal, multi-lineage differentiation, and ability to promote tissue regeneration, the last cell fraction or slowest to adhere to the collagen-coated surface (pre-plate 6; pp6) appears to be early, quiescent progenitor cells termed muscle-derived stem/progenitor cells (MDSPCs). The cell fractions preceding pp6 (pp1–5) are likely populations of more committed (differentiated) cells, including fibroblast- and myoblast-like cells. This technique may be used to isolate MDSPCs from skeletal muscle of humans or mice regardless of age, sex or disease state, although the yield of MDSPCs varies with age and health. MDSPCs can be used for regeneration of a variety of tissues including bone, articular cartilage, skeletal and cardiac muscle, and nerve. MDSPCs are currently being tested in clinical trials for treatment of urinary incontinence and myocardial infarction. MDSPCs from young mice have also been demonstrated to extend life span and healthspan in mouse models of accelerated aging through an apparent paracrine/endocrine mechanism. Here we detail methods for isolation and characterization of MDSPCs.

*Mitra Lavasani and Aiping Lu contributed equally.

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Acknowledgements

This work was supported by multiple institutes throughout many years including the National Institutes of Health, the US Department of Defense, the Muscular Dystrophy Association, the University of Pittsburgh Cancer Institute, and generous support from the William F. and Jean W. Donaldson Endowed Chair at the Children’s Hospital of Pittsburgh of UPMC, and the Henry J. Mankin Endowed Chair for Orthopaedic Research at the University of Pittsburgh.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Mitra Lavasani & Aiping Lu

  2. Stem Cell Research Center, University of Pittsburgh, Pittsburgh, PA, USA

    Mitra Lavasani, Aiping Lu, Seth D. Thompson & Johnny Huard

  3. Departments of Microbiology & Molecular Genetics and Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Paul D. Robbins & Johnny Huard

  4. University of Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, PA, USA

    Paul D. Robbins & Laura J. Niedernhofer

  5. Department of Microbiology and Molecular Genetics, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Laura J. Niedernhofer

Authors
  1. Mitra Lavasani
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  2. Aiping Lu
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  3. Seth D. Thompson
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  4. Paul D. Robbins
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  5. Johnny Huard
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  6. Laura J. Niedernhofer
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Correspondence to Laura J. Niedernhofer .

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Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Lavasani, M., Lu, A., Thompson, S.D., Robbins, P.D., Huard, J., Niedernhofer, L.J. (2013). Isolation of Muscle-Derived Stem/Progenitor Cells Based on Adhesion Characteristics to Collagen-Coated Surfaces. In: Turksen, K. (eds) Stem Cells and Aging. Methods in Molecular Biology, vol 976. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-317-6_5

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  • DOI: https://doi.org/10.1007/978-1-62703-317-6_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-316-9

  • Online ISBN: 978-1-62703-317-6

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