Direct Human Mitochondrial Transfer: A Novel Concept Based on the Endosymbiotic Theory
Direct Human Mitochondrial Transfer: A Novel Concept Based on the Endosymbiotic Theory
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DOI：
10.1016/j.transproceed.2013.11.133
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发表时间：
2014-05-01
影响因子：
0.9
通讯作者：
Gojo, S.
中科院分区：
文献类型：
--
作者：
Kitani, T.;Kami, D.;Gojo, S.
线粒体在真核生物中起着重要作用，线粒体功能障碍与多种疾病有关。因此，已经提出细胞间线粒体转移作为基于细胞的治疗的机制。此外，据报道，通过简单的共孵育使分离的线粒体细胞内化可改善受体细胞中的线粒体功能。然而，分离的线粒体内化的大量证据仍然缺乏，其确切的机制仍然难以捉摸。我们测试了是否可以通过简单的共孵育，使用荧光显微镜和流式细胞仪富集线粒体内化到培养的人类细胞。从子宫内膜腺源性间充质细胞（EMCs）或稳定表达子宫内膜靶向红色荧光蛋白（EMCs-DsRed-mito）的EMCs中分离线粒体，并通过抗线粒体抗体缀合的微珠富集。将它们与稳定表达绿色荧光蛋白（GFP）的同基因EMCs共孵育。活体荧光成像清楚地显示，DsRed标记的线粒体聚集在细胞核周围稳定表达GFP的EMCs的细胞质中。流式细胞术证实在受体细胞内存在GFP和DsRed双阳性细胞的不同群体。此外，转移效率依赖于线粒体浓度，表明人类细胞可能具有内在化线粒体的固有能力。因此，本研究支持应用同基因线粒体直接转移作为治疗线粒体功能障碍相关疾病的新方法。
Mitochondria play an essential role in eukaryotes, and mitochondrial dysfunction is implicated in several diseases. Therefore, intercellular mitochondrial transfer has been proposed as a mechanism for cell-based therapy. In addition, internalization of isolated mitochondria cells by simple coincubation was reported to improve mitochondrial function in the recipient cells. However, substantial evidence for internalization of isolated mitochondria is still lacking, and its precise mechanism remains elusive. We tested whether enriched mitochondria can be internalized into cultured human cells by simple coincubation using fluorescence microscopy and flow cytometry. Mitochondria were isolated from endometrial gland derived mesenchymal cells (EMCs) or EMCs stably expressing mitochondrial-targeted red fluorescent protein (EMCs-DsRed-mito), and enriched by anti-mitochondrial antibody-conjugated microbeads. They were coincubated with isogeneic EMCs stably expressing green fluorescent protein (GFP). Live fluorescence imaging clearly showed that DsRed-labeled mitochondria accumulated in the cytoplasm of EMCs stably expressing GFP around the nucleus. Flow cytometry confirmed the presence of a distinct population of GFP and DsRed double-positive cells within the recipient cells. In addition, transfer efficiency depended on mitochondrial concentration, indicating that human cells may possess the inherent ability to internalize mitochondria. Therefore, this study supports the application of direct transfer of isogeneic mitochondria as a novel approach for the treatment of diseases associated with mitochondrial dysfunction.