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首页> 外文期刊>Stem Cells >Characterization of a new NIH-registered variant human embryonic stem cell line, BG01V: a tool for human embryonic stem cell research.
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Characterization of a new NIH-registered variant human embryonic stem cell line, BG01V: a tool for human embryonic stem cell research.

机译：新型NIH注册的人类胚胎干细胞系BG01V的特性：人类胚胎干细胞研究的工具。

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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.
机译：人类胚胎干细胞（hESC）提供了可用于多种研究和基于细胞的疗法的多种细胞类型的可再生来源。表征这些细胞可提供有关其当前状态的重要信息，并为后续操作提供相关细节。例如，鉴定培养过程中表达的基因，以及传代后的时间表达顺序以及影响所有三个胚层形成的条件，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。尽管几种hESC品系在延长的培养过程中已显示出核型不稳定性，但某些选择的品系表现出与其正常亲本品系相似的特征。这样的变异株系可能是极好的工具，并且对于不关心染色体数目的中试研究和体外分化研究而言，它们是细胞的丰富来源，类似于目前胚胎癌细胞系所起的作用。至关重要的是，应在体外广泛繁殖，扩增和操纵过程中以遗传和蛋白质组学水平对细胞进行调查。在这里，我们描述了变体hESC系BG01V的全面表征，该变体源自核型正常的亲代hESC系BG01。我们的表征过程采用细胞遗传学分析，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，甲基化分析以及免疫表型和畸胎瘤形成，以及筛选。用于细菌，真菌，支原体和人类病原体的污染。

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