Genetic manipulation by means of microcell-mediated transfer of normal human chromosomes into recipient mouse cells.
Genetic manipulation by means of microcell-mediated transfer of normal human chromosomes into recipient mouse cells.
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通过微细胞介导将正常人类染色体转移到受体小鼠细胞中进行遗传操作。
DOI：
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发表时间：
1980
影响因子：
11.1
通讯作者：
R. Brown
中科院分区：
文献类型：
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作者：
C. A. Mcneill;R. Brown
微细胞介导的染色体转移是一种生产核型简单杂交种的创新方法。这种基因转移方法，利用长时间的秋碱处理形成的微核，已用于啮齿动物系统。由于无法获得来自二倍体人类细胞的大量微核群体，将这项技术扩展到包括正常人类遗传物质的转移一直受到阻碍。本报告描述了40-60%的正常人类成纤维细胞产生微核。这些微核细胞通过离心和细胞松弛素B联合处理去核，得到的微细胞被纯化并融合到受体小鼠（LMTK-）细胞中。包含单个人类染色体的微细胞杂交克隆已从三个单独的融合实验中分离出来。从融合到核型分析，这些杂交种的产生时间为6周。随着大约2 × 10（-6）的转移频率，单个完整的人类染色体已成为小鼠基因组的一个功能元素。
Microcell-mediated chromosome transfer is an innovative approach to the production of karyotypically simple hybrids. This method of gene transfer, employing micronuclei formed by prolonged Colcemid treatment, has been utilized for rodent systems. Expansion of this technology to include transfer of normal human genetic material has been hindered because large micronucleate populations from diploid human cells have been unobtainable. This report describes the production of micronuclei in 40-60% of normal human fibroblasts. These micronucleated cells have been enucleated by combining centrifugation and cytochalasin B treatment, and the resultant microcells have been purified and fused to recipient mouse (LMTK-) cells. Microcell hybrid clones containing a single human chromosome have been isolated from three separate fusion experiments. The time course for production of these hybrids, from fusion to karyotypic analysis, was 6 weeks. With a transfer frequency of about 2 x 10(-6), a single intact human chromosome has become a functioning element of the murine genome.