Efficient long-term cryopreservation of pluripotent stem cells at -80 °C.
Efficient long-term cryopreservation of pluripotent stem cells at -80 °C.
复制标题
-80°C 高效长期冷冻保存多能干细胞。
DOI：
10.1038/srep34476
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发表时间：
2016
影响因子：
4.6
通讯作者：
Han,Xu
中科院分区：
文献类型：
--
作者：
Yuan,Ye;Yang,Ying;Tian,Yuchen;Park,Jinkyu;Dai,Aihua;Roberts,RMichael;Liu,Yang;Han,Xu
Current long term cryopreservation of cell stocks routinely requires the use of liquid nitrogen (LN2), because commonly used cryopreservation media containing cell membrane permeating cryoprotectants are thermally unstable when frozen at higher storage temperatures, e.g. −80 °C. This instability leads to ice recrystallization, causing progressive loss of cell viability over time under the storage conditions provided by most laboratory deep freezers. The dependency on LN2for cell storage significantly increases operational expense and raises issues related to impaired working efficiency and safety. Here we demonstrate that addition of Ficoll 70 to cryoprotectant solutions significantly improves system thermal stability at the working temperature (~−80 °C) of laboratory deep freezers. Moreover, a medium comprised of Ficoll 70 and dimethyl sulfoxide (DMSO) in presence or absence of fetal bovine serum (FBS) can provide reliable cryopreservation of various kinds of human and porcine pluripotent stem cells at −80 °C for periods that extend up to at least one year, with the post-thaw viability, plating efficiency, and full retention of pluripotent phenotype comparable to that achieved with LN2storage. These results illustrate the practicability of a promising long-term cryopreservation method that completely eliminates the need for LN2.
