Chromosome transfer in mature oocytes.
Chromosome transfer in mature oocytes.
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成熟卵母细胞中的染色体转移。
DOI：
10.1038/nprot.2010.75
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发表时间：
2010-06
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
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作者：
在这篇文章中，我们描述了详细的协议之间的分离和转移成熟，中期II-逮捕卵母细胞的纺锤体染色体复合物。简而言之，使用偏光显微镜观察纺锤体-染色体复合体，并将其提取到膜封闭的核质体中。然后通过核质体-胞质体膜融合将染色体重新引入来自另一雌性的去核受体卵（胞质体）中。新重建的卵母细胞由来自一个雌性的核遗传物质和来自另一个雌性的细胞质组分（包括线粒体和线粒体DNA（mtDNA））组成。这种方法产生适合受精和产生胚胎干细胞或健康后代的有发育能力的卵母细胞。该方案最初是为猴卵母细胞开发的，但也可用于其他物种，包括小鼠和人类卵母细胞。潜在的临床应用包括线粒体基因替代疗法，以防止线粒体DNA突变的传播和治疗卵母细胞胞质缺陷引起的不孕症。从两个雌性分离的卵母细胞群之间的染色体转移可以在2小时内完成。
In this article, we describe detailed protocols for the isolation and transfer of spindle–chromosomal complexes between mature, metaphase II-arrested oocytes. In brief, the spindle–chromosomal complex is visualized using a polarized microscope and extracted into a membrane-enclosed karyoplast. Chromosomes are then reintroduced into an enucleated recipient egg (cytoplast), derived from another female, by karyoplast–cytoplast membrane fusion. Newly reconstructed oocytes consist of nuclear genetic material from one female and cytoplasmic components, including mitochondria and mitochondrial DNA (mtDNA), from another female. This approach yields developmentally competent oocytes suitable for fertilization and producing embryonic stem cells or healthy offspring. The protocol was initially developed for monkey oocytes but can also be used in other species, including mouse and human oocytes. Potential clinical applications include mitochondrial gene replacement therapy to prevent transmission of mtDNA mutations and treatment of infertility caused by cytoplasmic defects in oocytes. Chromosome transfer between the cohorts of oocytes isolated from two females can be completed within 2 h.