Ambry Genetics, Aliso Viejo, CA 92656, USA.
Mol Biotechnol. 2010 Nov;46(3):234-42. doi: 10.1007/s12033-010-9294-1.
Human embryonic and induced pluripotent stem cells (ESCs, iPSCs) that are cultured for an extended period of time are susceptible to genomic instability. Chromosomal aberrations decrease the reliability and reproducibility of experiments and could deem the cells unusable for therapeutic purposes. The genetic stability of human ESCs and iPSCs is commonly monitored by karyotype analysis. However, this low-resolution technique can only identify large aneuploidies. A reliable, high-resolution technique to detect genomic aberrations at a cost comparable to karyotyping is needed to better characterize stem cell lines. We have designed a stem cell focused array-comparative genomic hybridization microarray that covers the entire genome at high resolution with increased probe coverage in over 60 stem cell associated genes and more than 195 cancer related genes. Several iPSC lines were analyzed using the focused microarray and compared with either karyotyping or a standard Agilent 44K microarray. In addition to the abnormalities detected by these platforms, the custom microarray identified several small duplications spanning stem cell and/or cancer related genes. Scientists using a stem cell focused microarray to characterize their stem cells will be aware of the structural variants present in their cells and be more confident in their experimental results.
经过长时间培养的人类胚胎干细胞（ESCs）和诱导多能干细胞（iPSCs）容易发生基因组不稳定性。染色体畸变降低了实验的可靠性和可重复性，并可能使细胞无法用于治疗目的。人类 ESCs 和 iPSCs 的遗传稳定性通常通过核型分析来监测。然而，这种低分辨率技术只能识别大的非整倍体。需要一种可靠的、高分辨率的技术来检测基因组畸变，其成本应与核型分析相当，以便更好地表征干细胞系。我们设计了一种针对干细胞的比较基因组杂交微阵列，该微阵列以高分辨率覆盖整个基因组，在 60 多个与干细胞相关的基因和 195 多个与癌症相关的基因中增加了探针覆盖率。使用聚焦微阵列分析了几个 iPSC 系，并与核型分析或标准的安捷伦 44K 微阵列进行了比较。除了这些平台检测到的异常外，定制微阵列还鉴定了几个跨越干细胞和/或癌症相关基因的小重复。使用针对干细胞的微阵列来表征其干细胞的科学家将能够了解其细胞中存在的结构变异，并对其实验结果更有信心。
