INSERM Unité Mixte de Recherche S972, Université Paris-Sud, Unité Mixte de Recherche S972, and Département Hospitalo-Universitaire Hepatinov, Paul Brousse Hospital, Villejuif, France; INSERM Unité Mixte de Recherche S986, Institut Fédératif de Recherche 93, Bicêtre Hospital, Kremlin-Bicêtre, France; Department of Cytogenetics, INSERM U935, Béclère Hospital, Clamart, France; Centros de Investigación Biomédica en Red de Diabetes y Obesidad, Centro de Investigación Principe Felipe, Eduardo Primo Yúfera 3, Valencia, Spain; Molecular Genetics Center, Centre National de la Recherche Scientifique, Unité Propre de Recherche 3404, Gif-sur-Yvette, Université Paris-Sud, Orsay, France.
INSERM Unité Mixte de Recherche S972, Université Paris-Sud, Unité Mixte de Recherche S972, and Département Hospitalo-Universitaire Hepatinov, Paul Brousse Hospital, Villejuif, France; INSERM Unité Mixte de Recherche S986, Institut Fédératif de Recherche 93, Bicêtre Hospital, Kremlin-Bicêtre, France; Department of Cytogenetics, INSERM U935, Béclère Hospital, Clamart, France; Centros de Investigación Biomédica en Red de Diabetes y Obesidad, Centro de Investigación Principe Felipe, Eduardo Primo Yúfera 3, Valencia, Spain; Molecular Genetics Center, Centre National de la Recherche Scientifique, Unité Propre de Recherche 3404, Gif-sur-Yvette, Université Paris-Sud, Orsay, France
Stem Cells Transl Med. 2014 Jun;3(6):686-91. doi: 10.5966/sctm.2013-0158. Epub 2014 Apr 15.
The use of synthetic messenger RNAs to generate human induced pluripotent stem cells (iPSCs) is particularly appealing for potential regenerative medicine applications, because it overcomes the common drawbacks of DNA-based or virus-based reprogramming strategies, including transgene integration in particular. We compared the genomic integrity of mRNA-derived iPSCs with that of retrovirus-derived iPSCs generated in strictly comparable conditions, by single-nucleotide polymorphism (SNP) and copy number variation (CNV) analyses. We showed that mRNA-derived iPSCs do not differ significantly from the parental fibroblasts in SNP analysis, whereas retrovirus-derived iPSCs do. We found that the number of CNVs seemed independent of the reprogramming method, instead appearing to be clone-dependent. Furthermore, differentiation studies indicated that mRNA-derived iPSCs differentiated efficiently into hepatoblasts and that these cells did not load additional CNVs during differentiation. The integration-free hepatoblasts that were generated constitute a new tool for the study of diseased hepatocytes derived from patients' iPSCs and their use in the context of stem cell-derived hepatocyte transplantation. Our findings also highlight the need to conduct careful studies on genome integrity for the selection of iPSC lines before using them for further applications.
利用合成信使 RNA 来生成人类诱导多能干细胞（iPSC）对于潜在的再生医学应用特别有吸引力，因为它克服了基于 DNA 或基于病毒的重编程策略的常见缺点，特别是转基因的整合。我们通过单核苷酸多态性（SNP）和拷贝数变异（CNV）分析，比较了在严格可比条件下生成的 mRNA 衍生的 iPSC 与逆转录病毒衍生的 iPSC 的基因组完整性。我们表明，mRNA 衍生的 iPSC 在 SNP 分析中与亲本成纤维细胞没有显著差异，而逆转录病毒衍生的 iPSC 则有差异。我们发现，CNV 的数量似乎与重编程方法无关，而是与克隆有关。此外，分化研究表明，mRNA 衍生的 iPSC 能有效地分化为肝母细胞，并且这些细胞在分化过程中不会加载额外的 CNV。生成的无整合的肝母细胞为研究源自患者 iPSC 的患病肝细胞以及在干细胞衍生的肝细胞移植背景下使用它们提供了一种新工具。我们的研究结果还强调了在进一步应用之前，需要对 iPSC 系的基因组完整性进行仔细研究，以便进行选择。
