Simultaneous identification of chromatid replication and of human chromosomes in metaphases of man-mouse somatic cell hybrids. (With 1 color plate).
Simultaneous identification of chromatid replication and of human chromosomes in metaphases of man-mouse somatic cell hybrids. (With 1 color plate).
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在人-小鼠体细胞杂交中期同时鉴定染色单体复制和人类染色体。
DOI：
10.1159/000130814
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发表时间：
1977
期刊：
影响因子：
--
通讯作者：
M. Siniscalco
中科院分区：
文献类型：
--
作者：
B. Alhadeff;M. Velivasakis;M. Siniscalco
我们实验室正在进行的一项研究项目旨在调查早期人鼠体细胞杂交细胞中人类染色体与小鼠染色体不同步复制的理论可能性。为了验证这一假设，我们制定了以下方案，该方案允许在同一中期同时鉴定染色单体复制和人类染色体。该协议本质上是FPG （PLRRY和WOLFF， 1974）和Giemsa-1 I （BOBROW和CROSS， 1974）两种标准技术的结合。第一种方法可以识别染色单体交换，而第二种方法可以区分由不稳定的人鼠杂交细胞保留的人类染色体。混合细胞”。从我们的细胞库中获得的人鼠杂交体细胞克隆用于测试下面描述的程序。这些杂交克隆是在我们实验室通过仙台病毒诱导的L-A9小鼠细胞（LITTLEFIELD, 1964）与人类二倍体成纤维母细胞融合在人类基因定位的其他研究过程中产生的（MILLER等人，1971；SERAVALLI等人，1975）。根据细胞遗传学和生物化学分析的标准技术，对这些杂种的人类染色体和人类遗传标记的含量进行了充分的鉴定。当在标准MEM中生长时，补充15”/o ECS。这些杂交细胞在大约20-22小时内完成了它们的循环。杂交细胞的原始培养物通常用标准培养基保存在25厘米的falcon t ‘l ’培养皿中。一天
An on-going research project in our laboratory aims at investigating the theoretical possibility that human chromosomes in early man-mouse somatic hybrid cells may not replicate synchronously with those of the mouse. In order to test this hypothesis, we worked out the following protocol, which permits the simultaneous identification of chromatid repli cation and of the human chromosomes in the same metaphase. This protocol is essentially a combination of two standard techniques known under the terms FPG (PLRRY and WOLFF, 1974) and Giemsa-1 I (BOBROW and CROSS, 1974). The first one allows the identification of chromatid exchanges, while the second differentiates human chromosomes retained by the unstable man-mouse hybrid cells.Hybrid Cells'. Man-mouse hybrid somatic cell clones from our cell library were used to test the procedure described below. These hybrid clones had been produced in our laboratory by Sendai virusinduced fusion of L-A9 murine cells (LITTLEFIELD, 1964) with human diploid fibro blasts during the course of other studies on human gene mapping (MILLER ct al „1971; SERAVALLI et al „1975). These hybrids had been fully characterized for their content of human chromosomes and of human genetic markers according to standard techniques for cytogenetic and biochemical analyses. When grown in standard MEM supplemented with 15"/o ECS. these hybrid cells complete their cycle in about 20-22 h. Combined FPG-Giemsa-ll technique. Stock cultures of hybrid cells were normally maintained in 25-cm-Falcon T-l'lasks with standard medium. The day