IRMB, Univ Montpellier, INSERM, CHU Montpellier, Montpellier, France.
CHU Montpellier, Department of Cell and Tissue Engineering, Hospital Saint-Eloi, Montpellier, France.
Stem Cells. 2018 Jun;36(6):814-821. doi: 10.1002/stem.2797. Epub 2018 Feb 18.
Human induced pluripotent stem cells (hiPSCs) have the potential to differentiate virtually into any cell type in unlimited quantities. Therefore, they are ideal for in vitro tissue modeling or to produce cells for clinical use. Importantly, and differently from immortalized and cancer cell lines, the hiPSC genome scrupulously reproduces that of the cell from which they were derived. However, hiPSCs can develop genetic abnormalities during reprogramming or prolonged cell culture, such as aneuploidies or oncogenic mutations (e.g., in TP53). Therefore, hiPSC genome integrity must be routinely monitored because serious genome alterations would greatly compromise their usefulness or safety of use. Here, we reviewed hiPSC genome quality control monitoring methods and laboratory practice. Indeed, due to their frequency and functional consequences, recurrent genetic defects found in cultured hiPSCs are inacceptable and their appearance should be monitored by routine screening. Hence, for research purposes, we propose that the genome of hiPSC lines should be systematically screened at derivation, at least by karyotyping, and then regularly (every 12 weeks) during experiments, for instance with polymerase chain reaction-based techniques. For some specific applications, such as research on aging, cell cycle, apoptosis or cancer, other tests (e.g., TP53 mutation detection) should also be included. For clinical use, in addition to karyotyping, we advise exome sequencing. Stem Cells 2018;36:814-821.
人类诱导多能干细胞（hiPSCs）具有潜在的能力，可以无限量地分化为几乎任何类型的细胞。因此，它们非常适合体外组织建模或产生用于临床应用的细胞。重要的是，与永生化细胞系和癌细胞系不同，hiPSC 基因组严格复制其来源细胞的基因组。然而，hiPSCs 在重编程或长期细胞培养过程中可能会发生遗传异常，如非整倍体或致癌突变（例如，TP53 中的突变）。因此，必须定期监测 hiPSC 基因组完整性，因为严重的基因组改变会极大地影响其有用性或使用安全性。在这里，我们回顾了 hiPSC 基因组质量控制监测方法和实验室实践。事实上，由于它们的频率和功能后果，在培养的 hiPSCs 中发现的反复出现的遗传缺陷是不可接受的，其出现应该通过常规筛选进行监测。因此，对于研究目的，我们建议在衍生时系统地筛选 hiPSC 系的基因组，至少通过核型分析进行筛选，然后在实验过程中定期（例如每 12 周）进行筛选，例如使用聚合酶链反应（PCR）技术。对于某些特定的应用，如衰老、细胞周期、细胞凋亡或癌症的研究，还应包括其他测试（例如，TP53 突变检测）。对于临床用途，除了核型分析外，我们还建议进行外显子组测序。《干细胞》2018 年；36:814-821。
