无线粒体DNA的ρ°206细胞的培养及其呼吸链功能损害

Culture of human cell lines lacking mitochondrial DNA and its mitochondrial respiratory chain defects.

培养无线粒体DNA的 ρ°2 0 6细胞 ,观察其生长特性 ,应用电镜化学方法及检测MTT代谢率观察线粒体活性。结果发现 ρ°2 0 6细胞具有嘧啶依赖性 ,血清中微量的嘧啶即可支持其生长增殖成克隆。在选择培养基中 ρ°2 0 6细胞第 5天大量死亡 ,第 12天左右完全死亡 ,台盘蓝染色证实选择培养第 6天以后已无活细胞存在。ρ°2 0 6细胞的MTT代谢率仅为 143B·TK-细胞的 2 0 %左右 ,细胞色素氧化酶电镜细胞化学从形态学角度首次证实ρ°2 0 6细胞无细胞色素氧化酶活性 ,其线粒体数量及结构无明显异常改变。上述观察在转线粒体研究中具有一定的应用价值。

The study was designed to investigate the growth features of human cell lines lacking mitochondrial DNA(ρ°206 cells)and its mitochondrial activity assessed by electron microscopic cytochemistry and determination of MTT metabolism.The results showed that low levels of pyrimidines present in sera permited limited growth of ρ°206 cells,that ρ°206 cells died largely at 5th day in selective midium and died completely around 12 days after selective culture,there was no cells alive at 6 days after selective culture confirmed by trypan blue exclusion The MTT metabolism of ρ 0206 cells was only around 20% of that of 143B TK - cells. There was no cytichrome oxidase(COX) activity in mitochondria of ρ 0206 cells confirmed by COX electron microscopic cytochemistry. No abnormal changes of mitochondrial numbers and structures were found. These observations provided an applicable information in the transmitochondrial study.