Screening of human pluripotent stem cells using CGH and FISH reveals low-grade mosaic aneuploidy and a recurrent amplification of chromosome 1q
Screening of human pluripotent stem cells using CGH and FISH reveals low-grade mosaic aneuploidy and a recurrent amplification of chromosome 1q
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DOI：
10.1038/ejhg.2012.128
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发表时间：
2012-12-01
影响因子：
5.2
通讯作者：
Rienstein, Shlomit
中科院分区：
文献类型：
--
作者：
Dekel-Naftali, Michal;Aviram-Goldring, Ayala;Rienstein, Shlomit
人类胚胎干细胞(HESCs)的多能性和增殖能力使其成为基础和应用研究以及治疗医学的良好来源。人类诱导多能细胞(HiPSCs)的引入为患者量身定制的再生医学疗法带来了巨大的希望。然而，要将hESCs和hiPSCs用于治疗目的，必须保持培养中的长期基因组稳定性。直到最近，G显带分析一直被认为是检测干细胞染色体异常的默认方法。我们的目标是应用荧光原位杂交(FISH)和比较基因组杂交(CGH)来筛选多能干细胞，这将使我们能够更好地识别干细胞基因组中的染色体异常。我们对三个hESC系和两个hPSC系进行了长期培养研究。用FISH探针(12，13，16，17，18，21，X，Y)检测不同代次的非整倍体率。在hESCs和hiPSCs的早期传代中，基因组完整性被证明是保持的，但在传代后期，我们在hESCs中观察到低比例的嵌合体，这意味着传代数量和增加的非整倍体率之间存在直接的相关性。此外，CGH分析显示了一种反复出现的基因组不稳定，涉及到染色体1Q的获得。这一发现在两个不同来源的无关细胞系中被检测到，这意味着染色体1Q的获得可能在培养中赋予克隆优势。这些发现只能被传统的细胞遗传学方法部分检测到，强调了使用分子细胞遗传学方法追踪干细胞基因组不稳定性的重要性。《欧洲人类遗传学杂志》(2012年)2012481255；doi：10.1038/ejhg.2012.128；2012年6月20日在线发布
Pluripotency and proliferative capacity of human embryonic stem cells (hESCs) make them a promising source for basic and applied research as well as in therapeutic medicine. The introduction of human induced pluripotent cells (hiPSCs) holds great promise for patient-tailored regenerative medicine therapies. However, for hESCs and hiPSCs to be applied for therapeutic purposes, long-term genomic stability in culture must be maintained. Until recently, G-banding analysis was considered as the default approach for detecting chromosomal abnormalities in stem cells. Our goal in this study was to apply fluorescence in-situ hybridization (FISH) and comparative genomic hybridization (CGH) for the screening of pluripotent stem cells, which will enable us identifying chromosomal abnormalities in stem cells genome with a better resolution. We studied three hESC lines and two hiPSC lines over long-term culture. Aneuploidy rates were evaluated at different passages, using FISH probes (12,13,16,17,18,21,X,Y). Genomic integrity was shown to be maintained at early passages of hESCs and hiPSCs but, at late passages, we observed low rates mosaiciam in hESCs, which implies a direct correlation between number of passages and increased aneuploidy rate. In addition, CGH analysis revealed a recurrent genomic instability, involving the gain of chromosome 1q. This finding was detected in two unrelated cell lines of different origin and implies that gains of chromosome 1q may endow a clonal advantage in culture. These findings, which could only partially be detected by conventional cytogenetic methods, emphasize the importance of using molecular cytogenetic methods for tracking genomic instability in stem cells. European Journal of Human Genetics (2012) 20, 1248-1255; doi:10.1038/ejhg.2012.128; published online 20 June 2012