Sarafian Raquel, Morato-Marques Mariana, Borsoi Juliana, Pereira Lygia Veiga
National Laboratory for Embryonic Stem Cells (LaNCE), Department of Genetics and Evolutionary Biology, Biosciences Institute, University of São Paulo, SP 05508-900, Brazil.
National Laboratory for Embryonic Stem Cells (LaNCE), Department of Genetics and Evolutionary Biology, Biosciences Institute, University of São Paulo, SP 05508-900, Brazil.
Stem Cell Res. 2018 Apr;28:66-70. doi: 10.1016/j.scr.2018.01.030. Epub 2018 Jan 31.
The ability to reprogram somatic cells into induced pluripotent stem cells (hiPSCs) has led to the generation of large collections of cell lines from thousands of individuals with specific phenotypes, many of which will be shared among different research groups as invaluable tools for biomedical research. As hiPSC-based research involves extensive culture of many cell lines, the issue periodic cell line identification is particularly important to ensure that cell line identity remains accurate. Here we analyzed the different commercially available genotyping methods considering ease of in-house genotyping, cost and informativeness, and applied one of them in our workflow for hiPSC generation. We show that the chosen STR method was able to establish a unique DNA profile for each of the 35 individuals/hiPSC lines at the examined sites, as well as identify two discrepancies resulting from inadvertently exchanged samples. Our results highlight the importance of hiPSC line genotyping by an in-house method that allows periodic cell line identification and demonstrate that STR is a useful approach to supplement less frequent karyotyping and epigenetic evaluations.
将体细胞重编程为诱导多能干细胞（hiPSC）的能力，已促使人们从数千名具有特定表型的个体中生成了大量细胞系，其中许多细胞系将作为生物医学研究的宝贵工具在不同研究小组之间共享。由于基于hiPSC的研究涉及许多细胞系的广泛培养，因此定期进行细胞系鉴定对于确保细胞系身份准确尤为重要。在这里，我们考虑到内部基因分型的难易程度、成本和信息量，分析了不同的商业可用基因分型方法，并将其中一种应用于我们生成hiPSC的工作流程中。我们表明，所选择的STR方法能够在所检测的位点为35个个体/hiPSC系中的每一个建立独特的DNA图谱，还能识别出因样本意外交换而导致的两个差异。我们的结果强调了通过一种允许定期进行细胞系鉴定的内部方法对hiPSC系进行基因分型的重要性，并证明STR是一种补充不太频繁的核型分析和表观遗传评估的有用方法。
