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[通过胚胎干细胞与体细胞融合获得的杂交细胞中同源染色体区域复制的荧光原位杂交分析]

[FISH-analysis of regional replication in homologous chromosomes in hybrid cells obtained by fusion of embryonic stem cells with somatic cells].

作者信息

Podriadchikova O L, Pristiazhniuk I E, Matveeva N M, Serov O L

出版信息

Tsitologiia. 2009;51(6):500-5.

Abstract

The paper concerns FISH-analysis of regional replication of parental chromosomes 1, 3 and 6 in hybrid cells obtained by fusion between Mus musculus embryonic stem cells (ESC) and M. caroli splenocytes. The data demonstrated that parental chromosomes in the hybrid cells with near-diploid karyotype showed synchronous replication in 70-75% of tested cells that was comparable with diploid ESC and diploid fibroblasts. Synchronous replication of parental chromosomes in hybrid cells with near-triploid karyotype was observed in 46-57% of tested cells. However, it was correct in the case of hybrid cells with three copies of the tested chromosomes whereas the ratio of synchronous replication in triploid cells with two copies was comparable or similar to that in diploid cells. Hybrid cells with near-tetraploid karyotype showed high ratio of asynchronous replication (over 50%) comparable to those tetraploid ESC and tetraploid fibroblasts but significantly distinguished from diploid cells. Thus, most hybrid cells with two copies of tested parental chromosomes showed their synchronous replication. Unfortunately, the FISH-analysis is poor informative when it is used for studying cells with more than two copies of tested chromosomes.

摘要

本文涉及对小家鼠胚胎干细胞(ESC)与卡罗小鼠脾细胞融合获得的杂交细胞中亲代1号、3号和6号染色体区域复制的荧光原位杂交(FISH)分析。数据表明,具有近二倍体核型的杂交细胞中的亲代染色体在70%-75%的测试细胞中显示出同步复制,这与二倍体ESC和二倍体成纤维细胞相当。在具有近三倍体核型的杂交细胞中,46%-57%的测试细胞观察到亲代染色体的同步复制。然而,对于具有三份测试染色体的杂交细胞情况是正确的,而具有两份测试染色体的三倍体细胞中的同步复制比例与二倍体细胞中的相当或相似。具有近四倍体核型的杂交细胞显示出高比例的异步复制(超过50%),与四倍体ESC和四倍体成纤维细胞相当,但与二倍体细胞有显著区别。因此,大多数具有两份测试亲代染色体的杂交细胞显示出同步复制。不幸的是,当FISH分析用于研究具有两份以上测试染色体的细胞时,其信息量不足。

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