title: Assaying Human Myogenic Progenitor Cell Activity by Reconstitution of Muscle Fibers and Satellite Cells in Immunodeficient Mice.
abstract: Comparing the functional myogenic potential of various human cell populations is an important step in the preclinical evaluation of cell transplantation as a means to treat human muscle disease and degeneration. Culture systems allow one to gage the potential of cell populations to proliferate and undergo myogenic differentiation under specific conditions. An in vivo assay evaluates the ability of cells to differentiate and generate muscle fibers within a natural environment, and importantly, evaluates the potential of donor cells to reconstitute the satellite cell niche. In this chapter, we describe a technique for isolating mononuclear cells from human muscle samples, and a method of xenotransplantation for assessing functional myogenic potential in vivo. Briefly, cell populations are injected into the pre-irradiated and regenerating muscle of immunodeficient mice. The injected muscle is frozen at specific time points after injection and cryosections analyzed by immunostaining. The number of human dystrophin-expressing fibers and the number of Pax7(+) human lamin A/C(+) nuclei are determined, which provides a quantitative method of comparing the in vivo functional potential of cell populations.
authors: Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N, Seattle, WA, 98109, USA. mparker@fredhutch.org.|Department of Medicine, University of Washington, 356420, Seattle, WA, 98195, USA. mparker@fredhutch.org.
authors: Maura H
authors: MH
authors: Parker
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D006085:Graft Survival; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology*; D012038:Regeneration; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology*; D055153:Stem Cell Niche; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
keywords: Human muscle; Immunodeficient mice; Muscle fibers; Muscle progenitors; Regeneration; Satellite cells; Xenotransplant