Platelet-Rich Plasma Promotes the Proliferation of Human Muscle Derived Progenitor Cells and Maintains Their Stemness

Abstract

Human muscle-derived progenitor cells (hMDPCs) offer great promise for muscle cell-based regenerative medicine; however, prolonged ex-vivo expansion using animal sera is necessary to acquire sufficient cells for transplantation. Due to the risks associated with the use of animal sera, the development of a strategy for the ex vivo expansion of hMDPCs is required. The purpose of this study was to investigate the efficacy of using platelet-rich plasma (PRP) for the ex-vivo expansion of hMDPCs. Pre-plated MDPCs, myoendothelial cells, and pericytes are three populations of hMDPCs that we isolated by the modified pre-plate technique and Fluorescence Activated Cell Sorting (FACS), respectively. Pooled allogeneic human PRP was obtained from a local blood bank, and the effect that thrombin-activated PRP-releasate supplemented media had on the ex-vivo expansion of the hMDPCs was tested against FBS supplemented media, both in vitro and in vivo. PRP significantly enhanced short and long-term cell proliferation, with or without FBS supplementation. Antibody-neutralization of PDGF significantly blocked the mitogenic/proliferative effects that PRP had on the hMDPCs. A more stable and sustained expression of markers associated with stemness, and a decreased expression of lineage specific markers was observed in the PRP-expanded cells when compared with the FBS-expanded cells. The in vitro osteogenic, chondrogenic, and myogenic differentiation capacities of the hMDPCs were not altered when expanded in media supplemented with PRP. All populations of hMDPCs that were expanded in PRP supplemented media retained their ability to regenerate myofibers in vivo. Our data demonstrated that PRP promoted the proliferation and maintained the multi-differentiation capacities of the hMDPCs during ex-vivo expansion by maintaining the cells in an undifferentiated state. Moreover, PDGF appears to be a key contributing factor to the beneficial effect that PRP has on the proliferation of hMDPCs.

Platelet-Rich Blood Derivatives for Stem Cell-Based Tissue Engineering and Regeneration

“…Interestingly, PRP was shown to up-regulate the pluripotency markers and decrease lineage specific marker expression in human MSCs, human muscle derived progenitor cells, and adipose derived stem cells [44, 75]. Moreover, PRP has been demonstrated to protect cell viability during cryopreservation [43].…”

Physiology and metabolism of tissue-engineered skeletal muscle

“…14 For example, human muscle-derived progenitor cells isolated by Huard and colleagues can differentiate into skeletal muscle cells, cardiomyocytes, endothelial cells, pericytes, chondrocytes and bone cells. 15, 16 A fraction of these cells with elevated levels of aldehyde dehydrogenase resist oxidative stress exhibit increased proliferation and ability to form myotubes, 17 suggesting a promising cell source for muscle tissue engineering. Embryonic stem cells and induced pluripotent stem cells are another cell source that can differentiate into muscle cells and hold the promise of providing a ready source of satellite cells and myoblasts, 18–20 but further work is needed to produce sufficient numbers of cells generate contractile myofibers, and address potential safety concerns.…”

Autologous platelet rich plasma enhances satellite cells expression of MyoD and exerts angiogenic and antifibrotic effects in experimental rat model of traumatic skeletal muscle injury

“…PRP also had a potent effect on the proliferation of human muscle-derived progenitor cells (hMDPCs) and keep their stemness. In addition, it maintained their multi-lineage differentiation ability which gives the cells a vast facility to differentiate into osteogenic and chondrogenic line in vitro and a myogenic line both in vitro and in vivo [65] .…”

“…In mature adult muscle, TGF β1 negatively affects skeletal muscle regeneration by inhibiting satellite cell proliferation and myofiber fusion. Furthermore, in abnormal muscle regeneration there is a persistent inflammatory response and over expression of TGF β1 and myostatin which stimulate the transformation of myogenic cells into myofibroblasts with the formation of fibrotic tissue to replace damaged myofibers [65,73] . Accordingly, reducing the levels of TGF-β1has been proven to be beneficial for several myopathic conditions [74][75][76][77] .…”