Transferring isolated mitochondria into tissue culture cells.
Transferring isolated mitochondria into tissue culture cells.
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DOI：
10.1093/nar/gks639
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发表时间：
2012-10
影响因子：
14.9
通讯作者：
Koob MD
中科院分区：
文献类型：
--
作者：
Yang YW;Koob MD
我们已经开发了一种新的方法，将大量分离的线粒体导入组织培养细胞。已经发现将线粒体直接显微注射到典型的哺乳动物细胞中是不切实际的，这是由于线粒体相对于显微注射针的尺寸大。为了解决这个问题，我们通过适当大小的显微注射针将分离的线粒体注射到啮齿动物卵母细胞或单细胞胚胎中，这些卵母细胞或单细胞胚胎比组织培养细胞大得多，然后使用修改后的保持针取出含有注射的线粒体的“线粒体”细胞片段。然后通过病毒介导的膜融合将这些有丝分裂质体与受体细胞融合，并将注射的线粒体转移到组织培养细胞的细胞质中。由于小鼠卵母细胞含有大量的小鼠线粒体，这些线粒体与注射的线粒体一起沿着重新填充受体小鼠细胞，因此我们使用沙鼠单细胞胚胎或大鼠卵母细胞包装注射的小鼠线粒体。我们发现，沙鼠线粒体DNA（mtDNA）是不保持在受体rho0小鼠细胞和大鼠mtDNA最初复制，但很快就完全取代了注射的小鼠mtDNA，因此与这两个程序小鼠细胞同质的小鼠mtDNA在注射的线粒体。
We have developed a new method for introducing large numbers of isolated mitochondria into tissue culture cells. Direct microinjection of mitochondria into typical mammalian cells has been found to be impractical due to the large size of mitochondria relative to microinjection needles. To circumvent this problem, we inject isolated mitochondria through appropriately sized microinjection needles into rodent oocytes or single-cell embryos, which are much larger than tissue culture cells, and then withdraw a ‘mitocytoplast’ cell fragment containing the injected mitochondria using a modified holding needle. These mitocytoplasts are then fused to recipient cells through viral-mediated membrane fusion and the injected mitochondria are transferred into the cytoplasm of the tissue culture cell. Since mouse oocytes contain large numbers of mouse mitochondria that repopulate recipient mouse cells along with the injected mitochondria, we used either gerbil single-cell embryos or rat oocytes to package injected mouse mitochondria. We found that the gerbil mitochondrial DNA (mtDNA) is not maintained in recipient rho0 mouse cells and that rat mtDNA initially replicated but was soon completely replaced by the injected mouse mtDNA, and so with both procedures mouse cells homoplasmic for the mouse mtDNA in the injected mitochondria were obtained.
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影响因子：
64.5
作者：
KING, MP;ATTARDI, G
通讯作者：
ATTARDI, G
DOI：
10.1073/pnas.71.5.1681
发表时间：
1974-01-01
影响因子：
11.1
作者：
BUNN, CL;WALLACE, DC;EISENSTA.JM
通讯作者：
EISENSTA.JM
与人线粒体3302a> g突变相关的分子功能障碍（mt-trnaleu（uur））基因中的分子功能障碍。
影响因子：
14.9
作者：
Maniura-Weber, Katharina;Helm, Mark;Engemann, Katrin;Eckertz, Sabrina;Moellers, Myriam;Schauen, Matthias;Hayrapetyan, Armine;von Kleist-Retzow, Juergen-Christoph;Lightowlers, Robert N.;Bindoff, Laurence A.;Wiesner, Rudolf J.
通讯作者：
Wiesner, Rudolf J.
影响因子：
3
作者：
Pinkert, CA;Irwin, MH;Moffatt, RJ
通讯作者：
Moffatt, RJ
DOI：
10.1006/bbrc.2001.4646
发表时间：
2001-04-06
影响因子：
3.1
作者：
Yamaoka, M;Mikami, T;Hayashi, JI
通讯作者：
Hayashi, JI