Human embryonic stem cell lines isolation, cultivation, and characterization
Human embryonic stem cell lines isolation, cultivation, and characterization
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DOI：
10.1007/s11626-010-9282-6
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发表时间：
2010-04-01
影响因子：
2.1
通讯作者：
Kiselev, Sergei L.
中科院分区：
文献类型：
--
作者：
Lagarkova, Maria A.;Eremeev, Artem V.;Kiselev, Sergei L.
自 1998 年建立第一个 hESC 系以来，全世界已衍生出大量人类胚胎干细胞 (hESC) 系。尽管 hESC 系具有许多共同特征，其中最重要的是多能性，但 hESC 系在转录谱、遗传和表观遗传状态方面存在显着差异。这些差异可能源于细胞系的个体遗传学以及其处理（例如分离和培养）的变化。为了尽量减少后者的差异，应执行标准化的培养方案和实验室间综合研究。在本报告中，我们总结了 hESC 系的衍生和表征以及 hESC 适应新培养方案的经验。我们已成功衍生出 5 种 hESC 系，并根据先前制定的标准对它们进行了表征，包括特定标记的表达以及体外和体内分化的能力。其中四个系，即 hESM01-04，最初是使用小鼠成纤维细胞作为饲养层衍生的，目前使用 mTeSR1 和 Matrigel 在无饲养层、无血清条件下维持。第五株系 hESMK05 是使用 mTeSR1 和 Matrigel 在无饲养层、无血清条件下衍生的。细胞系在 70 多次传代中仍保持其多能状态和正常核型，可供科学界使用。
A large number of human embryonic stem cell (hESC) lines have been derived worldwide since the first hESC line establishment in 1998. Despite many common characteristics, most important of which is the pluripotency, hESC lines vary significantly in their transcriptional profiles, genetic, and epigenetic state. These differences may arise both from individual genetics of the cell lines and from variations in their handling such as isolation and cultivation. In order to minimize the latter differences, the standardized protocols of cultivation and inter-laboratory comprehensive studies should be performed. In this report, we summarized our experience of derivation and characterization of hESC lines as well as of adaptation of hESCs to novel cultivation protocols. We have successfully derived five hESC lines and characterized them by previously established criteria, including expression of specific markers and the capacity to differentiate both in vitro and in vivo. Four of these lines, namely hESM01-04, were initially derived using mouse fibroblasts as a feeder and currently are maintained under feeder-free, serum-free conditions using mTeSR1 and Matrigel. The fifth line, hESMK05 was derived in feeder-free, serum-free conditions using mTeSR1 and Matrigel. Cell lines retain their pluripotent status and normal karyotype for more than 70 passages and are available to the scientific community.