Chromosome number variation in three mouse embryonic stem cell lines during culture
Chromosome number variation in three mouse embryonic stem cell lines during culture
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DOI：
10.1007/s10616-008-9164-x
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发表时间：
2008-09-01
期刊：
影响因子：
2.2
通讯作者：
Garagna, Silvia
中科院分区：
文献类型：
--
作者：
Rebuzzini, Paola;Neri, Tui;Garagna, Silvia
小鼠胚胎干细胞（mESC）自1981年建立以来，一直缺乏对培养过程中染色体变化的系统研究。在这项研究中，我们报告的细胞遗传学分析的结果进行了三个mESC线（命名为UPV 02，UPV 06和UPV 08）培养3个月。在时间间隔上，染色体数目的变化与未分化状态的标志物的表达一起，即，测定OCT-4、SSEA-1、FOM-1和碱性磷酸酶活性。在3个月的培养期间，三个mESC系显示整倍体中期细胞的进行性损失。在分析的最新传代中，染色体异常累积。在整个培养过程中，中着丝粒染色体是最常见的染色体异常。有趣的是，在整倍性下降的同时，或在整倍性下降后的几次传代中，碱性磷酸酶活性部分或全部丧失，而OCT-4、SSEA-1和FOM-1茎标记在整个培养期间始终呈阳性。我们的研究结果表明，有必要在培养过程中进行核型分析，以开发新的培养条件，以保持正确的染色体互补，在长期培养的mESC系。
Although mouse embryonic stem cell lines (mESCs) have been established since 1981, systematic studies about chromosomal changes during culture are lacking. In this study, we report the results of a cytogenetic analysis performed on three mESC lines (named UPV02, UPV06 and UPV08) cultured for a period of 3 months. At time intervals, the variation of the chromosome number together with the expression of markers of the undifferentiated status, i.e., OCT-4, SSEA-1, FOM-1 and alkaline phosphatase activity, were determined. The three mESC lines showed a progressive loss of euploid metaphases during the 3 months period of culture. Chromosome abnormalities were accumulated at the latest passages analysed. Metacentric chromosomes were the most frequent chromosome abnormality observed throughout the period of culture. Interestingly, in coincidence with, or few passages after, the drop of euploidy, the alkaline phosphatase activity was partially or totally lost, whereas the OCT-4, SSEA-1 and FOM-1 stem markers were always positive throughout the period of culture. Our results remark the necessity to perform the karyotype analysis during culture in order to develop new culture conditions to maintain the correct chromosome complement in long-term culture of mESC lines.