Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage
Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage
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DOI：
10.1016/j.bbrc.2006.04.135
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发表时间：
2006-07-07
影响因子：
3.1
通讯作者：
Nakatsuji, Norio
中科院分区：
文献类型：
--
作者：
Suemori, Hirofumi;Yasuchika, Kentaro;Nakatsuji, Norio
人类胚胎干细胞(HES)细胞系被认为是医学研究以及在细胞治疗和药物开发中应用的宝贵资源。然而，为了实现对HES细胞的这种利用，涉及到HES细胞使用的方案，例如用于建立、繁殖和冷冻保存的方案，仍然需要改进。在这里，我们报告了一种建立HES细胞系的有效方法及其详细的特性。此外，我们开发了一种新的批量传代技术，当HES细胞系在培养条件下保持长达2年的核型完整性时，该技术可以保持其核型完整性。最后，我们证明了一种简化的玻璃化冷冻保存技术在细胞存活率方面大大优于标准的缓慢降温方法。这些结果提供了有价值的信息，有助于实现将HES细胞应用于疾病治疗所需的大规模HES细胞培养的目标。(C)2006 Elsevier Inc.保留所有权利。
Human ES (hES) cell lines are considered to be a valuable resource for medical research and for applications in cell therapy and drug discovery. For such utilization of hES cells to be realized, however, protocols involved in the use of hES cells, such as those for establishment, propagation, and cryopreservation, have still to be improved. Here, we report on an efficient method for the establishment of hES cell lines and its detailed characterization. Additionally, we developed a new bulk-passaging technique that preserves the karyotypic integrity of hES cell lines when maintained in culture for up to 2 years. Finally, we show that a simplified vitrification cryopreservation technique is vastly superior to standard slow-cooling methods with respect to cell viability. These results provide valuable information that will assist in achieving the goal of the large-scale hES cell culture required for the application of hES cells to disease therapy. (c) 2006 Elsevier Inc. All rights reserved.