Mitochondria transfer enhances proliferation, migration, and osteogenic differentiation of bone marrow mesenchymal stem cell and promotes bone defect healing
Mitochondria transfer enhances proliferation, migration, and osteogenic differentiation of bone marrow mesenchymal stem cell and promotes bone defect healing
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线粒体移植增强骨髓间充质干细胞的增殖、迁移和成骨分化，促进骨缺损愈合
DOI：
10.1186/s13287-020-01704-9
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发表时间：
2020-06-25
影响因子：
7.5
通讯作者：
Deng, Xuliang
中科院分区：
文献类型：
--
作者：
Guo, Yusi;Chi, Xiaopei;Deng, Xuliang
背景骨髓间充质干细胞（BMSC）移植被认为是一种有前途的骨缺损修复方法。然而，在移植过程中，骨髓间充质干细胞的功能和活力可能会受到损害，由于体外培养时间延长，老化，和患者的疾病状况。受自然发生在受伤组织内的线粒体自发转移，以挽救细胞或组织功能的启发，我们研究了人工线粒体转移到移植前BMSCs在体外是否可以提高细胞功能，提高其对骨缺损修复situ.MethodsMitochondria的治疗效果，从供体BMSCs中分离，并转移到受体BMSCs的同一批次和通道。随后，通过活细胞成像、Cell Counting Kit-8测定、细胞周期分析、Ki 67染色、c-Myc表达的qPCR和Western印迹分析以及β-半乳糖苷酶染色评价增殖能力和细胞衰老的变化。通过transwell迁移试验、伤口划痕愈合和细胞运动试验评价迁移能力。采用碱性磷酸酶（ALP）染色、茜素红染色、qPCR和Western blot检测Runx 2和BMP 2的表达，观察线粒体转染对BMSCs体外成骨能力的影响。在此之后，体内实验进行移植到大鼠颅骨临界大小的骨缺损模型的骨髓间充质干细胞。术后4周和8周行显微CT扫描和组织学观察，评价原位成骨情况。最后，为了建立细胞的行为变化和有氧代谢之间的相关性，OXPHOS（氧化磷酸化）和ATP的生产进行了评估和抑制有氧呼吸寡霉素performed. ResultsMitochondrial-recipient骨髓间充质干细胞表现出显着增强的增殖和迁移，并增加成骨诱导成骨。体内实验结果显示，自体骨髓基质干细胞原位移植后，新骨形成增多。增加OXPHOS活性和ATP的产生，观察到，寡霉素抑制后衰减的增殖，迁移和成骨分化诱导的线粒体transfer. ConclusionsMitochondrial转移的增强是一种可行的技术，以提高BMSC功能在体外，促进骨缺损修复原位通过上调有氧代谢。结果表明，线粒体转移可能是优化干细胞治疗功能的一种新的有前途的技术。
BackgroundBone marrow-derived mesenchymal stem cell (BMSC) transplantation is considered a promising therapeutic approach for bone defect repair. However, during the transplantation procedure, the functions and viability of BMSCs may be impaired due to extended durations of in vitro culture, aging, and disease conditions of patients. Inspired by spontaneous intercellular mitochondria transfer that naturally occurs within injured tissues to rescue cellular or tissue function, we investigated whether artificial mitochondria transfer into pre-transplant BMSCs in vitro could improve cellular function and enhance their therapeutic effects on bone defect repair in situ.MethodsMitochondria were isolated from donor BMSCs and transferred into recipient BMSCs of the same batch and passage. Subsequently, changes in proliferative capacity and cell senescence were evaluated by live cell imaging, Cell Counting Kit-8 assay, cell cycle analysis, Ki67 staining, qPCR and Western blot analysis of c-Myc expression, and β-galactosidase staining. Migration ability was evaluated by the transwell migration assay, wound scratch healing, and cell motility tests. Alkaline phosphatase (ALP) staining, Alizarin Red staining, and combined with qPCR and Western blot analyses of Runx2 and BMP2 were performed to elucidate the effects of mitochondria transfer on the osteogenic potential of BMSCs in vitro. After that, in vivo experiments were performed by transplanting mitochondria-recipient BMSCs into a rat cranial critical-size bone defect model. Micro CT scanning and histological analysis were conducted at 4 and 8 weeks after transplantation to evaluate osteogenesis in situ. Finally, in order to establish the correlation between cellular behavioral changes and aerobic metabolism, OXPHOS (oxidative phosphorylation) and ATP production were assessed and inhibition of aerobic respiration by oligomycin was performed.ResultsMitochondria-recipient BMSCs exhibited significantly enhanced proliferation and migration, and increased osteogenesis upon osteogenic induction. The in vivo results showed more new bone formation after transplantation of mitochondria-recipient BMSCs in situ. Increased OXPHOS activity and ATP production were observed, which upon inhibition by oligomycin attenuated the enhancement of proliferation, migration, and osteogenic differentiation induced by mitochondria transfer.ConclusionsMitochondria transfer is a feasible technique to enhance BMSC function in vitro and promote bone defect repair in situ through the upregulation of aerobic metabolism. The results indicated that mitochondria transfer may be a novel promising technique for optimizing stem cell therapeutic function.