Murine and Human Myogenic Cells Identified by Elevated Aldehyde Dehydrogenase Activity: Implications for Muscle Regeneration and Repair

Abstract

BackgroundDespite the initial promise of myoblast transfer therapy to restore dystrophin in Duchenne muscular dystrophy patients, clinical efficacy has been limited, primarily by poor cell survival post-transplantation. Murine muscle derived stem cells (MDSCs) isolated from slowly adhering cells (SACs) via the preplate technique, induce greater muscle regeneration than murine myoblasts, primarily due to improved post-transplantation survival, which is conferred by their increased stress resistance capacity. Aldehyde dehydrogenase (ALDH) represents a family of enzymes with important morphogenic as well as oxidative damage mitigating roles and has been found to be a marker of stem cells in both normal and malignant tissue. In this study, we hypothesized that elevated ALDH levels could identify murine and human muscle derived cell (hMDC) progenitors, endowed with enhanced stress resistance and muscle regeneration capacity.Methodology/Principal FindingsSkeletal muscle progenitors were isolated from murine and human skeletal muscle by a modified preplate technique and unfractionated enzymatic digestion, respectively. ALDHhi subpopulations isolated by fluorescence activate cell sorting demonstrated increased proliferation and myogenic differentiation capacities compared to their ALDHlo counterparts when cultivated in oxidative and inflammatory stress media conditions. This behavior correlated with increased intracellular levels of reduced glutathione and superoxide dismutase. ALDHhi murine myoblasts were observed to exhibit an increased muscle regenerative potential compared to ALDHlo myoblasts, undergo multipotent differentiation (osteogenic and chondrogenic), and were found predominately in the SAC fraction, characteristics that are also observed in murine MDSCs. Likewise, human ALDHhi hMDCs demonstrated superior muscle regenerative capacity compared to ALDHlo hMDCs.ConclusionsThe methodology of isolating myogenic cells on the basis of elevated ALDH activity yielded cells with increased stress resistance, a behavior that conferred increased regenerative capacity of dystrophic murine skeletal muscle. This result demonstrates the critical role of stress resistance in myogenic cell therapy as well as confirms the role of ALDH as a marker for rapid isolation of murine and human myogenic progenitors for cell therapy.

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

“…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). Their high therapeutic value is likely due to their superior survival capability under conditions of oxidative and hypoxic stresses and high expression of antioxidants relative to more differentiated cells, such as myoblasts (47,48). Most recently, our findings showed that i.p.…”

Lying low but ready for action: the quiescent muscle satellite cell

“…In muscle, two populations of satellite cells have been characterized on the basis of their levels of aldehyde dehydrogenase activity. Satellite cells with high levels of activity of this enzyme were shown to be more resistant to cell death in vitro when exposed to H 2 O 2 and in vivo after cell transplantation [92], thus providing a criterion for myogenic progenitors with increased value as therapeutic agents for treating dystrophic muscle [93]. Similarly, high glutathione reductase and superoxide dismutase activity correlated with enhanced regenerative activity of muscle satellite cells [94].…”

Glutathione peroxidase 3, a new retinoid target gene, is crucial for human skeletal muscle precursor cell survival

“…Recently, we determined that high aldehyde dehydrogenase activity (ALDH high ) is associated with improved cell viability in human myoblasts (Jean et al, 2011). In addition, Vauchez et al and Vella et al have isolated ALDH high muscle progenitors cells that exhibit increased stress resistance and regenerative capacity (Vauchez et al, 2009;Vella et al, 2011). Several ALDH enzymes, including Aldh1a1, Aldh1a2 and Aldh1a3 in humans, catalyze the irreversible oxidation of vitamin A (VA) to retinoic acid (RA), which binds and activates nuclear retinoic acid receptor (RAR)/retinoid X receptor (RXR) heterodimers to regulate the transcription of target genes that are important for development, morphogenesis and differentiation (Jackson et al, 2011;Samarut and Rochette-Egly, 2012).…”