外源性线粒体DNA在细胞内的重建(简报)

REPOPULATION WITH EXOGENOUS MITOCHONDRIA IN THE CELL

近年来发现人类多种神经肌肉疾病存在线粒体电子传递链(electron transport chain,ETC)缺陷[1-7].由于线粒体在遗传上受核基因和线粒体基因双重控制,给确定ETC缺陷的来源造成困难.转线粒体DNA技术是线粒体同无线粒体DNA的细胞(ρ0cells)融合[8,9],形成转线粒体DNA细胞系(mtDNA-transferted cell line,也称cytoplasmic hy-brids,简称cybrids),使病人的线粒体DNA(mito-chondrial DNA,mtDNA)同正常人的mtDNA在完全一致的细胞背景中表达,排除细胞内其它因素的干扰,单独研究mtDNA对细胞功能的影响.

To create human mitochondrial DNA(mtDNA)-transferred cells as cell model of mitochondria defects-related diseases, platelet of normal young and old subjects were isolated as donor of mtDNA, then fused with mtDNA-depleted cells (ρ0 cells) under induction of PEG1500. Auxtrophy test, cytochrome c oxidase activity assay and PCR amplification of mtDNA were done to confirm the transferring of mtDNA. Cell clones were visible in the medium 10 to 15 days after fusion, which grew well in medium lacking uridine, had positive COX activity and contained objective fragment of mtDNA by PCR, opposite to ρ0 cells. Transferring of mtDNA to ρ0 cells was identified, and mtDNA-transferred cell models were successfully created.