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首页> 外文期刊>International Journal of Cell Biology >Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-TermCulture
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Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-TermCulture

机译：长期培养后人皮肤成纤维细胞X-失活状态和细胞遗传学稳定性的评价

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摘要

Human primary fibroblasts are a popular type of somatic cells for the production of induced pluripotent stem (iPS) cells. Here we characterized biological properties of primary fibroblasts in terms of cell-growth rate, cytogenetic stability, and the number of inactive X chromosomes during long-term passaging. We produced eight lines of female human dermal fibroblasts (HDFs) and found normal karyotype and expected pattern of X chromosome inactivation (XCI) at low passages (Passage P1-5). However, four out of the eight HDF lines at high passage numbers (≥ P10) exhibited duplicated hallmarks of inactive X chromosome including two punctuate signals of histone H3 lysine 27 trimethylation (H3K27me3) and X inactive-specific transcript (XIST) RNA signals in approximately 8.5–18.5% of the cells. Our data suggest that the copy number of inactive X chromosomes in a subset of female HDF is increased by a two-fold. Consistently, DNA fluorescent in situ hybridization (FISH) identified 3-4 copies of X chromosomes in one nucleus in this subset of cells with two inactive Xs. We conclude that female HDF cultures exhibit a higher risk of genetic anomalies such as carrying an increased number of X chromosomes including both active and inactive X chromosomes at a high passage (≥ P10).
机译：人类原代成纤维细胞是一种普遍的体细胞类型，用于产生诱导性多能干（iPS）细胞。在这里，我们根据细胞生长速率，细胞遗传学稳定性以及长期传代过程中非活性X染色体的数量来表征原代成纤维细胞的生物学特性。我们生产了八种雌性人类皮肤成纤维细胞（HDF），并在低传代（传代P1-5）下发现了正常的染色体核型和X染色体失活（XCI）的预期模式。但是，在高传代数（≥P10）的8个HDF品系中，有4个表现出无活性X染色体的重复特征，包括大约2个组蛋白H3赖氨酸27三甲基化（H3K27me3）的标点信号和X个非活性特异性转录本（XIST）RNA信号8.5–18.5％的细胞。我们的数据表明，女性HDF子集中非活动X染色体的拷贝数增加了两倍。一致地，DNA荧光原位杂交（FISH）在具有两个非活性Xs的这一细胞亚群中的一个核中鉴定了3-4个X染色体拷贝。我们得出的结论是，女性HDF培养物具有较高的遗传异常风险，例如携带高数量（≥P10）的X染色体（包括活动X染色体和非活动X染色体）数量增加。

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