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富含血小板的血浆促进人肌肉来源祖细胞的增殖并维持其干性。

Platelet-rich plasma promotes the proliferation of human muscle derived progenitor cells and maintains their stemness.

机构信息

Department of Orthopedic Surgery, Stem Cell Research Center, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.

出版信息

PLoS One. 2013 Jun 7;8(6):e64923. doi: 10.1371/journal.pone.0064923. Print 2013.

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.

摘要

人肌肉源祖细胞(hMDPCs)为基于肌细胞的再生医学提供了巨大的希望;然而,为了获得用于移植的足够细胞,需要使用动物血清进行长时间的体外扩增。由于使用动物血清存在风险,因此需要开发一种用于 hMDPCs 体外扩增的策略。本研究旨在探讨使用富含血小板的血浆(PRP)进行 hMDPCs 体外扩增的效果。我们通过改良的预培养技术和荧光激活细胞分选(FACS)分别分离了 hMDPCs 的三个群体:预培养 MDPCs、肌内皮细胞和周细胞。从当地血库获得了混合异体人 PRP,并测试了凝血酶激活的 PRP 释放物补充培养基对 hMDPCs 的体外和体内扩增效果,与 FBS 补充培养基进行了对比。PRP 显著增强了 hMDPCs 的短期和长期增殖,无论是否有 FBS 补充。PDGF 的抗体中和显著阻断了 PRP 对 hMDPCs 的有丝分裂/增殖作用。与 FBS 扩增细胞相比,PRP 扩增细胞观察到与干细胞相关的标记物的表达更加稳定和持续,谱系特异性标记物的表达降低。当在 PRP 补充的培养基中进行扩增时,hMDPCs 的体外成骨、软骨和成肌分化能力并未改变。在 PRP 补充培养基中扩增的所有 hMDPCs 群体在体内都保留了再生肌纤维的能力。我们的数据表明,PRP 通过使细胞保持未分化状态,促进了 hMDPCs 的增殖并维持了其在体外扩增过程中的多向分化能力。此外,PDGF 似乎是 PRP 促进 hMDPCs 增殖的有益作用的关键因素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/292f/3676442/5c34bf03062f/pone.0064923.g001.jpg

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