Isolation of a Highly Myogenic CD34-Negative Subset of Human Skeletal Muscle Cells Free of Adipogenic Potential

Abstract

The differentiation of multipotent cells into undesirable lineages is a significant risk factor when performing cell therapy. In muscular diseases, myofiber loss can be associated with progressive fat accumulation that is one of the primary factors leading to decline of muscular strength. Therefore, to avoid any contribution of injected multipotent cells to fat deposition, we have searched for a highly myogenic but nonadipogenic muscle-derived cell population. We show that the myogenic marker CD56, which is the gold standard for myoblast-based therapy, was unable to separate muscle cells into myogenic and adipogenic fractions. Conversely, using the stem cell marker CD34, we were able to sort two distinct populations, CD34(+) and CD34(-), which have been thoroughly characterized in vitro and in vivo using an immunodeficient Rag2(-/-)gamma(c) (-/-) mouse model of muscle regeneration with or without adipose deposition. Our results demonstrate that both populations have equivalent capacities for in vitro amplification. The CD34(+) cells and CD34(-) cells exhibit equivalent myogenic potential, but only the CD34(-) population fails to differentiate into adipocytes in vitro and in vivo after transplantation into regenerative fat muscle. These data indicate that the muscle-derived cells constitute a heterogeneous population of cells with various differentiation potentials. The simple CD34 sorting allows isolation of myogenic cells with no adipogenic potential and therefore could be of high interest for cell therapy when fat is accumulated in diseased muscle.

Age-related changes in the features of porcine adult stem cells isolated from adipose tissue and skeletal muscle

“…ϩ ) progenitors in human muscle (37,38). Altogether, these data further support the idea that these cell populations may display different differentiation potentials.…”

“…Therefore, the present study was undertaken to enrich our knowledge on the phenotype and differentiation potential of skeletal muscle stromal vascular (SV) cells compared with adipose tissue SV cells in postnatal growing and adult animals. This study was based on the expression of a variety of well-characterized cell surface markers such as CD11b and CD14 for immune cells (36), CD31 for endothelial cells, CD34 for hematopoietic stem cells and other cells including cells from the endothelium (39), CD45 for hematopoietic cells, CD56 for muscle precursors (38,39), and CD90 (Thy-1) for MSCs (31). In the current study, the pig was used not only for its important role in animal protein supply for human nutrition but also for its relevance as a suitable animal model for several human disorders (27,44). )]…”

Adipose tissue stem cells meet preadipocyte commitment: going back to the future

“…Similarly, exclusion of CD34+ cells in muscle regeneration studies inhibits ectopic adipose formation both in vitro and in vivo ( 190 ), indicating that CD34-cells are incapable of adipogenesis. Finally, adipocytes isolated from WAT are noted as CD34+ ( 106 ).…”

Role of Pericytes in Skeletal Muscle Regeneration and Fat Accumulation

“…However, even mouse models of such disorders as DMD or obesity rarely show adipocytes in skeletal muscle. We used a model of muscle fatty degeneration reported in the literature [10,25,26,[54][55][56][57][58][59][60][61]. Injecting glycerol in the muscle destabilizes cell membranes, promoting myofiber damage, significant fat deposits along the muscle, and cell death.…”