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

Abstract

Summary 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 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) appear 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 myoblastlike 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 lifespan and healthspan in mouse models of accelerated aging through an apparent paracrine/endocrine mechanism. Here we detail methods for isolation and characterization of MDSPCs.

Extracellular vesicles and aging

“…Similarly, MSCs derived from the bone marrow of patients with Hutchinson-Gilford Progeroid Syndrome, a disease of accelerated aging, are defective in their ability to differentiate (41). In addition, muscle derived stem/progenitor cells (MDSPC) are adversely affected in accelerated and natural aged mice, displaying loss of proliferation and ability to differentiate in culture (11,42). Importantly, this dysfunction was demonstrated to directly contribute to age-related degenerative changes since intraperitoneal injection of only 106 functional, young MDSPCs was sufficient to extend healthspan and lifespan in two different mouse models of accelerated aging (11).…”

Human muscle–derived stem/progenitor cells promote functional murine peripheral nerve regeneration

“…Our recent data suggest that MDSPCs, which can be isolated from human skeletal muscles (hMDSPCs) using the same technique (39), are likely mesenchymal stem cells of muscle origin and have the ability to undergo multilineage differentiation (51). In the present study, we examine the fate of hMDSPCs in controlled culture conditions and their potential for functional nerve repair.…”

“…Stem/progenitor cells isolated from murine and human skeletal muscles by various methods give rise to progeny cells with neuronal and glial phenotypes (12,(32)(33)(34)(35)(36). Populations of slowly adhering cells isolated from skeletal muscle via the modified preplate technique called muscle-derived stem/progenitor cells (MDSPCs) (37)(38)(39) are characterized by sustained self-renewal, long-term proliferation, and multipotent differentiation capacities (37,40,41). MDSPCs can engraft and stimulate the regeneration of skeletal and cardiac muscles, bone, articular cartilage, and replenish the bone marrow of lethally irradiated mice (37,40,(42)(43)(44)(45)(46).…”

Fundamentals and application of magnetic particles in cell isolation and enrichment: a review

“…One of the simplest ways to separate target cells from a heterogeneous cell population is to harness the unique differential adhesion profile of different cells in the heterogeneous suspension (Lavasani et al, 2013, Brown et al, 2008, Laugwitz et al, 2005). Cells can be placed in culture with a growth medium, either on native polystyrene or coated with cell adhesion biomolecules, and over a given period of time specific cell populations will adhere to culture substrates.…”