Characterization of a new NIH-Registered variant human embryonic stem cell line, BG01V: A tool for human embryonic stem cell research
Characterization of a new NIH-Registered variant human embryonic stem cell line, BG01V: A tool for human embryonic stem cell research
复制标题
DOI：
10.1634/stemcells.2005-0315
复制
发表时间：
2006-03-01
期刊：
影响因子：
5.2
通讯作者：
Auerbach, Jonathan M.
中科院分区：
文献类型：
--
作者：
Plaia, Todd W.;Josephson, Richard;Auerbach, Jonathan M.
人类胚胎干细胞（hESCs）为研究和基于细胞的治疗提供了广泛的可再生细胞类型。描述这些单元格提供了有关其当前状态的重要信息，并为后续操作提供了相关细节。例如，鉴定在培养过程中表达的基因，以及它们在传代后的时间表达顺序和影响所有三种生殖层形成的条件，可能有助于生产用于治疗I型糖尿病的功能性β胰岛细胞。虽然一些hESC细胞系在长时间培养中表现出核型不稳定性，但选择的变异细胞系表现出与其正常亲本细胞系相似的特征。这些变异细胞系可能是前期研究和体外分化研究的良好工具和丰富的细胞来源，在这些研究中染色体数量不受关注，类似于目前胚胎癌细胞系所起的作用。在体外广泛繁殖、扩增和操作过程中，对细胞进行遗传和蛋白质组学水平的调查是至关重要的。在这里，我们描述了变异hESC系BG01V的综合特性，它来源于核型正常的亲本hESC系BG01。我们的鉴定过程包括细胞遗传学分析、短串联重复序列和HLA分型、线粒体DNA测序、使用定量逆转录-聚合酶链反应和微阵列的基因表达分析、端粒酶活性评估、甲基化分析、免疫表型和畸胎瘤形成，以及细菌、真菌、支原体和人类病原体污染筛查。
Human embryonic stem cells (hESCs) offer a renewable source of a wide range of cell types for use in research and cell-based therapies. Characterizing these cells provides important information about their current state and affords relevant details for subsequent manipulations. For example, identifying genes expressed during culture, as well as their temporal expression order after passaging and conditions influencing the formation of all three germ layers may be helpful for the production of functional beta islet cells used in treating type I diabetes. Although several hESC lines have demonstrated karyotypic instability during extended time in culture, select variant lines exhibit characteristics similar to their normal parental lines. Such variant lines may be excellent tools and abundant sources of cells for pilot studies and in vitro differentiation research in which chromosome number is not a concern, similar to the role currently played by embryonal carcinoma cell lines. It is crucial that the cells be surveyed at a genetic and proteomic level during extensive propagation, expansion, and manipulation in vitro. Here we describe a comprehensive characterization of the variant hESC line BG01V, which was derived from the karyotypically normal, parental hESC line BG01. Our characterization process employs cytogenetic analysis, short tandem repeat and HLA typing, mitochondrial DNA sequencing, gene expression analysis using quantitative reverse transcription-polymerase chain reaction and microarray, assessment of telomerase activity, methylation analysis, and immunophenotyping and teratoma formation, in addition to screening for bacterial, fungal, mycoplasma, and human pathogen contamination.