部分性克隆人胚胎干细胞系中的遗传和表观遗传 X 染色体变异。
Genetic and epigenetic X-chromosome variations in a parthenogenetic human embryonic stem cell line.
机构信息
Guangzhou Key Laboratory of Reproductive and Genetics; Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510150, China.
出版信息
J Assist Reprod Genet. 2011 Apr;28(4):303-13. doi: 10.1007/s10815-010-9517-1. Epub 2010 Dec 15.
PURPOSE
To assess the genetic and epigenetic status of parthenogenetic human embryonic stem cells (phESCs).
METHODS
Cytogenetics, X chromosome inactivation (XCI) and gene expression patterns were analyzed in one phESC line (FY-phES-018) that was derived from our laboratory.
RESULTS
FY-phES-018 cells displayed the classical characteristics of normal hESCs. These cells had a 46, XX karyotype, and no inactive X chromosomes were observed before passage 20. After being cultured long term in vitro, some cells lost one X, and the proportion of cells with only one X gradually increased. At passage 35, almost all the cells displayed a 45, XO karyotype. Interestingly, at passage 45, the recovery of the X-chromosome was observed, and XCI became detectable; the mosaic ratio of 46, XX to 45, XO was 67:33. After passage 60, most cells displayed the 46, XX karyotype again with a mosaic ratio of 97:3. Some aberrant genomic imprinting was also observed in these cells.
CONCLUSIONS
The phESCs line FY-phES-018 is both genetically and epigenetically unstable; therefore, further research is needed before using these cells.
目的
评估体细胞核移植胚胎干细胞（phESC）的遗传和表观遗传状态。
方法
对我们实验室中衍生的一条 phESC 系（FY-phES-018）进行了细胞遗传学、X 染色体失活（XCI）和基因表达模式分析。
结果
FY-phES-018 细胞表现出正常 hESC 的典型特征。这些细胞具有 46,XX 核型，在传代 20 次之前未观察到无活性的 X 染色体。在体外长期培养后，一些细胞丢失了一个 X 染色体，只有一个 X 染色体的细胞比例逐渐增加。在传代 35 时，几乎所有细胞都表现出 45,XO 核型。有趣的是，在传代 45 时，观察到 X 染色体的恢复，并且可以检测到 XCI；46,XX 与 45,XO 的镶嵌比例为 67:33。在传代 60 后，大多数细胞再次表现出 46,XX 核型，镶嵌比例为 97:3。这些细胞中还观察到一些异常的基因组印迹。
结论
phESC 系 FY-phES-018 在遗传和表观遗传上均不稳定；因此，在使用这些细胞之前需要进一步研究。
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