Enhancement of cell recovery for dissociated human embryonic stem cells after cryopreservation.
Enhancement of cell recovery for dissociated human embryonic stem cells after cryopreservation.
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冷冻保存后解离的人类胚胎干细胞的细胞恢复增强。
DOI：
10.1002/btpr.358
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发表时间：
2010-05
影响因子：
2.9
通讯作者：
Cui, Zhanfeng
中科院分区：
文献类型：
--
作者：
Xu, Xia;Cowley, Sally;Flaim, Christopher J.;James, William;Seymour, Lenard W.;Cui, Zhanfeng
由于人类胚胎干细胞的广泛应用，建立有效的冻存和后续培养方案以提高细胞回收率是至关重要的。我们已经开发了一种新的方案，用于冷冻保存解离的hES细胞和随后的培养。我们考察了含有7.5%二甲基亚砜（DMSO）（v/v %）和2.5%聚乙二醇（PEG）（w/v %）的冷冻液新配方对冻存后hES细胞存活和恢复的影响，并进一步研究了Rho相关激酶（ROCK）抑制剂和p53抑制剂联合使用对后续培养中细胞恢复的作用。与传统的缓慢冷冻方法（使用10%DMSO作为冷冻液，然后在培养的第一天在ROCK抑制剂的存在下培养）相比，我们发现，通过新的冷冻液，hES细胞的回收率显著提高了约30%（P < 0.05）。此外，在解冻后培养的第一天，10 μM ROCK抑制剂（Y-27632）和1 μM pifithrin-μ的存在进一步显著提高了细胞回收率，无论是饲养层依赖性培养还是非饲养层依赖性培养，均提高了约20%（P < 0.05）。hES细胞在使用这种新的方案进行冷冻保存和随后的培养后保持其未分化状态。此外，该方案是用于处理大量hES细胞的可扩展的冷冻保存方法。© 2009年美国化学工程师学会生物技术。程序：2010
Due to widespread applications of human embryonic stem (hES) cells, it is essential to establish effective protocols for cryopreservation and subsequent culture of hES cells to improve cell recovery. We have developed a new protocol for cryopreservation of dissociated hES cells and subsequent culture. We examined the effects of new formula of freezing solution containing 7.5% dimethylsulfoxide (DMSO) (v/v %) and 2.5% polyethylene glycol (PEG) (w/v %) on cell survival and recovery of hES cells after cryopreservation, and further investigated the role of the combination of Rho-associated kinase (ROCK) inhibitor and p53 inhibitor on cell recovery during the subsequent culture. Compared with the conventional slow-freezing method which uses 10% DMSO as a freezing solution and then cultured in the presence of ROCK inhibitor at the first day of culture, we found out that hES cell recovery was significantly enhanced by around 30 % (P < 0.05) by the new freezing solution. Moreover, at the first day of post-thaw culture, the presence of 10 μM ROCK inhibitor (Y-27632) and 1 μM pifithrin-μ together further significantly improved cell recovery by around 20% (P < 0.05) either for feeder-dependent or feeder-independent culture. hES cells remained their undifferentiated status after using this novel protocol for cryopreservation and subsequent culture. Furthermore, this protocol is a scalable cryopreservation method for handling large quantities of hES cells. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010
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影响因子：
19.6
作者：
Hauet, T;Goujon, JM;Vandewalle, A
通讯作者：
Vandewalle, A
影响因子：
4.7
作者：
Luo, J;Borgens, R;Shi, R
通讯作者：
Shi, R
影响因子：
2.6
作者：
Heng, Boon Chin
通讯作者：
Heng, Boon Chin
人类胚胎干细胞的批量玻璃化冷冻。
影响因子：
6.1
作者：
Li, Tao;Zhou, Canquan;Zhuang, Guanglun
通讯作者：
Zhuang, Guanglun
影响因子：
4
作者：
Valbuena, Diana;Sanchez-Luengo, Silvia;Simon, Carlos
通讯作者：
Simon, Carlos