背景:本系统综述描述了使人和动物间充质干细胞(MSC)永生化的最常用方法。本研究遵循PRISMA的规则,并在PROSPERO国际系统审查机构审查委员会注册,编号协议代码:CRD42020202465。
方法:数据搜索系统化基于“间充质干细胞”和“永生化”。“出版物的搜索期为2000年至2022年,使用的数据库为SCOPUS,pubmed,科学直接。搜索策略确定了384篇文章:SCOPUS数据库中的229篇文章,84在PUBMED,和71在科学直接。经过标题和摘要筛选,还有285篇文章。根据纳入和排除标准,本综述纳入了39篇文章。
结果:在28篇文章中,从人类和11只动物中永生化MSC。最常用的永生化方法是病毒转染。最常见的永生化细胞类型是来自骨髓的MSC,人和动物MSCs永生化最常用的基因是hTERT(39.3%)和SV40T(54.5%),分别。
结论:另外,据观察,尽管不到一半的研究进行了致瘤性试验来验证永生化的MSC,其他化验,如qRT-PCR,在软琼脂中形成菌落,核型,FISH,和细胞增殖,在大多数研究中对不同的MSC细胞传代进行。
BACKGROUND: This systematic review describes the most common methodologies for immortalizing human and animal mesenchymal stem cells (MSCs). This study follows the rules of PRISMA and is registered in the Institutional Review Board of PROSPERO International of systematic reviews, numbered protocol code: CRD42020202465.
METHODS: The data search systematization was based on the words \"mesenchymal stem cell\" AND \"immortalization.\" The search period for publications was between 2000 and 2022, and the databases used were SCOPUS, PUBMED, and SCIENCE DIRECT. The search strategies identified 384 articles: 229 in the SCOPUS database, 84 in PUBMED, and 71 in SCIENCE DIRECT. After screening by titles and abstracts, 285 articles remained. This review included thirty-nine articles according to the inclusion and exclusion criteria.
RESULTS: In 28 articles, MSCs were immortalized from humans and 11 animals. The most used immortalization methodology was viral transfection. The most common immortalized cell type was the MSC from bone marrow, and the most used gene for immortalizing human and animal MSCs was hTERT (39.3%) and SV40T (54.5%), respectively.
CONCLUSIONS: Also, it was observed that although less than half of the studies performed tumorigenicity assays to validate the immortalized MSCs, other assays, such as qRT-PCR, colony formation in soft agar, karyotype, FISH, and cell proliferation, were performed in most studies on distinct MSC cell passages.