A simple and efficient cryopreservation method for primate embryonic stem cells
A simple and efficient cryopreservation method for primate embryonic stem cells
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DOI：
10.1387/ijdb.041852tf
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发表时间：
2004-12-01
影响因子：
0.7
通讯作者：
Suemori, H
中科院分区：
文献类型：
--
作者：
Fujioka, T;Yasuchika, K;Suemori, H
人类胚胎干细胞（ES细胞）具有分化为所有细胞类型的潜力。由于这些细胞可能能够为移植治疗提供无限的细胞来源，因此有必要建立可靠的方法来处理和操作它们，包括人ES细胞冷冻保存。在这里，我们报告了一个简单而有效的冷冻保存方法的灵长类动物ES细胞系使用玻璃化在传统的冷冻管的发展。使用标准慢速冷却方法，食蟹猴ES细胞系的冷冻保存效率约为0.4%，而人ES细胞系的冷冻保存效率几乎为0%。然而，灵长类动物ES细胞系通过本发明的玻璃化方法使用常规冷冻管成功地冷冻保存，猴ES细胞的存活率为约6.5%，人ES细胞的存活率为12.2%。玻璃化ES细胞解冻后迅速恢复，并表现出与非玻璃化细胞无法区分的形态。此外，它们保留了正常的核型，并在解冻后继续表达ES细胞标记。因此，我们的玻璃化ES细胞冷冻保存方法扩展了灵长类动物ES细胞用于各种研究和临床目的的效用。
Human embryonic stem (ES) cells have the potential to differentiate into all cell types. As these cells may be able to provide an unlimited cell source for transplantation therapies, it is necessary to establish reliable methods for their handling and manipulation, including human ES cell cryopreservation. Here, we report the development of a simple and efficient cryopreservation method for primate ES cell lines using vitrification in conventional cryovials. Using standard slow-rate cooling methods, the cryopreservation efficiency for cynomoigus monkey ES cell lines was approximately 0.4%, while that for a human ES cell line was virtually 0%. Primate ES cell lines, however, were successfully cryopreserved by the present vitrification method using conventional cryovials yielding a survival rate of about 6.5% for monkey ES cells and 12.2% for human ES cells. Vitrified ES cells quickly recovered after thawing and exhibited a morphology indistinguishable from non-vitrified cells. In addition, they retained a normal karyotype and continued to express ES cell markers after thawing. Thus, our vitrification ES cell cryopreservation method expands the utility of primate ES cells for various research and clinical purposes.