中国作者重要发表 - ROCK抑制剂延长了无血清/营养剂冷冻保存单人类胚胎干细胞的生存期



ROCK抑制剂延长了无血清/营养剂冷冻保存单人类胚胎干细胞的生存期





ROCK inhibitor improves survival of cryopreserved serum/feeder-free single human embryonic stem cells





Li X., Krawetz R., Liu S., Meng G., Rancourt D.E. 2009/5/29 18:39:42

【发表评论】



打印\| 推荐给好友向好友发送电子邮件收件人：（请填写电子邮箱地址，如果有多个邮箱请用逗号”,”分开。）发件人：（电子邮箱地址）内容：（可选）最多填写200字。


Human Reproduction, 2009, Volume 24, Issue 3




BACKGROUND: Efficient slow freezing protocols within serum-free and feeder-free culture systems are crucial for the clinical application of human embryonic stem (hES) cells. Frequently, however, hES cells must be cryopreserved as clumps when using conventional slow freezing protocols, leading to lower survival rates during freeze-thaw and limiting their recovery and growth efficiency after thawing, as well as limiting downstream applications that require single cell suspensions. We describe a novel method to increase freeze-thaw survival and proliferation rate of single hES cells in serum-free and feeder-free culture conditions. METHODS: hES cells maintained on Matrigel-coated dishes were dissociated into single cells with Accutase and slow freezing. After thawing at 37°C, cells were cultured in mTeSR medium supplemented with 10 μM of Rho-associated kinase inhibitor Y-27632 for 1 day. RESULTS: The use of Y-27632 and Accutase significantly increases the survival of single hES cells after thawing compared with a control group (P < 0.01). Furthermore, by treatment of hES cell aggregates with EGTA to disrupt cell-cell interaction, we show that Y-27632 treatment does not directly affect hES cell apoptosis. Even in the presence of Y-27632, hES cells deficient in cell-cell interaction undergo apoptosis. Y-27632-treated freeze-thawed hES cells retain typical morphology, stable karyotype, expression of pluripotency markers and the potential to differentiate into derivatives of all three germ layers after long-term culture. CONCLUSIONS: The method described here allows for cryopreservation of single hES cells in serum-free and feeder-free conditions and therefore we believe this method will be ideal for current and future hES cell applications that are targeted towards a therapeutic end-point. © The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.



Correspondence Address: Rancourt, D. E.; Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB T2N 4N1, Canada; email: rancourt@ucalgary.ca



评论



请登录后发表评论，点击此处登录。

全部学科

疾病资源中心

高血压	慢性阻塞性肺病
脂肪性肝病	糖尿病
卒中	肺癌
冠心病	子宫内膜异位症
结直肠癌	乳腺癌
心力衰竭	心律失常

病例分析

甲状腺功能亢进合并黄疸

王燕燕王曙

上海交通大学附属瑞金医院内分泌科

患者，女，69岁。2009年1月无明显诱因下出现乏力，当时程度较轻，未予以重视。2009年3月患者乏力症状加重，尿色逐渐加深，大便习惯改变，颜色变淡。4月18日入我院感染科治疗，诉轻度头晕、心慌，体重减轻10kg。无肝区疼痛，无发热，无腹痛、腹泻、腹胀、里急后重，无恶性、呕吐等。入院半月前于外院就诊，查肝功能：ALT 601IU/L，AST 785IU/L，TBIL 97.7umol/L，白蛋白 41g/L，甲状腺功能：游离T3 30.6pmol/L，游离T4 51.9pmol/L，心电图示快速房颤。

医学数据库