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[胚胎干细胞杂交细胞中亲代染色体的可见和“隐蔽”分离]

[Visible and "cryptic" segregation of parental chromosomes in embryonic stem hybrid cells].

作者信息

Pristiazhniuk I E, Temirova S A, Menzorov A G, Kruglova A A, Matveeva N M, Serov O L

出版信息

Ontogenez. 2005 Mar-Apr;36(2):151-8.

Abstract

Chromosome segregation of the parental chromosomes was studied in 20 interspecific hybrid clones obtained by fusion of Mus musculus embryonic stem cells with Mus caroli splenocytes. FISH analysis with labeled species specific probes and microsatellite markers was used for identification of the parental chromosomes. Cytogenetic analysis has shown significant intra- and interclonal variability in chromosome numbers and ratios of the parental chromosomes in the hybrid cells: six clones contained all M. caroli chromosomes, nine clones showed moderate segregation of M. caroli chromosomes (from 1 to 7), and five clones showed extensive loss of M. caroli chromosomes (from 12 to complete loss of all M. caroli autosomes). Both methods demonstrated "cryptic" segregation of the somatic partner chromosomes. For instance, five clones with near-tetraploid chromosome sets contained only few M. caroli chromosomes (from 1 to 8). The data obtained suggest that the tetraploid chromosome set per se is not a sufficient criterion for conclusion on the absence of chromosome loss in the hybrid cells. Note that "cryptic" chromosome segregation occurred at a high frequency in the examined hybrid clones. Thus, "cryptic" segregation should be borne in mind for assessing pluripotency and genome reprogramming of embryonic stem hybrid cells.

摘要

通过小家鼠胚胎干细胞与斯氏小家鼠脾细胞融合获得了20个种间杂交克隆,并对亲代染色体的染色体分离进行了研究。使用标记的物种特异性探针和微卫星标记进行荧光原位杂交(FISH)分析,以鉴定亲代染色体。细胞遗传学分析表明,杂交细胞中染色体数目和亲代染色体比例存在显著的克隆内和克隆间变异性:六个克隆包含所有斯氏小家鼠染色体,九个克隆显示斯氏小家鼠染色体的中度分离(从1条到7条),五个克隆显示斯氏小家鼠染色体大量丢失(从12条到所有斯氏小家鼠常染色体完全丢失)。两种方法均显示了体细胞伙伴染色体的“隐匿性”分离。例如,五个具有近四倍体染色体组的克隆仅包含少数斯氏小家鼠染色体(从1条到8条)。获得的数据表明,四倍体染色体组本身并不是判断杂交细胞中不存在染色体丢失的充分标准。请注意,在所检查的杂交克隆中,“隐匿性”染色体分离发生的频率很高。因此,在评估胚胎干细胞杂交细胞的多能性和基因组重编程时应牢记“隐匿性”分离。

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