期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1980
卷号：77
期号：9
页码：5394-5398
DOI：10.1073/pnas.77.9.5394
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：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.
