Barriers in contribution of human mesenchymal stem cells to murine muscle regeneration

• 作者: Anabel S de la Garza-Rodea Antoine AF de Vries Cheryl Dambrot Dirk W van Bekkum Hester Boersma Shoshan Knaan-Shanzer
• 来源期刊: 世界实验医学杂志
• 学科: 医学
• 关键词: SKELETAL MUSCLE MUSCLE regeneration MUSCLE IMPLANTS MESENCHYMAL stem CELLS Satellite CELLS
• 年，卷（期）: 2015,（2）
• 页码范围: 140-153
• 页数: 14页
• 分类号: R

摘要

AIM:To study regeneration of damaged human and murine muscle implants and the contribution of added xenogeneic mesenchymal stem cells(MSCs).METHODS:Minced human or mouse skeletal muscle tissues were implanted together with human or mouse MSCs subcutaneously on the back of non-obese diabetic/severe combined immunodeficient mice.The muscle tissues(both human and murine)were minced with scalpels into small pieces(<1 mm3)and aliquoted in portions of 200 mm3.These portions were either cryopreserved in 10%dimethylsulfoxide or freshly implanted.Syngeneic or xenogeneic MSCs were added to the minced muscles directly before implantation.Implants were collected at 7,14,30 or 45 d after transplantation and processed for(immuno)histological analysis.The progression of muscle regeneration was assessed using a standard histological staining(hematoxylin-phloxinsaffron).Antibodies recognizing Pax7 and von Willebrand factor were used to detect the presence of satellite cells and blood vessels,respectively.To enable detection of the bone marrow-derived MSCs or their derivatives we used MSCs previously transduced with lentiviral vectors expressing a cytoplasmic LacZ gene.X-gal staining of the fixed tissues was used to detectβ-galactosidase-positive cells and myofibers.RESULTS:Myoregeneration in implants of fresh murine muscle was evident as early as day 7,and progressed with time to occupy 50%to 70%of the implants.Regeneration of fresh human muscle was slower.These observations of fresh muscle implants were in contrast to the regeneration of cryopreserved murine muscle that proceeded similarly to that of fresh tissue except for day 45(P<0.05).Cryopreserved human muscle showed minimal regeneration,suggesting that the freezing procedure was detrimental to human satellite cells.In fresh and cryopreserved mouse muscle supplemented with LacZ-tagged mouse MSCs,β-galactosidase-positive myofibers were identified early after grafting at the wellvascularized periphery of the implants.The contribution of human MSCs to murine myofiber formati