| 人胚胎干细胞程序降温保存的实验研究 |
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| 引用本文： | 杨鹏飞,丛笑倩,程启康,华泽钊,吴春芳,曹谊林.人胚胎干细胞程序降温保存的实验研究[J].实验生物学报,2005,38(3):247-256. |
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| 作者姓名： | 杨鹏飞 丛笑倩 程启康 华泽钊 吴春芳 曹谊林 |
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| 作者单位： | [1]上海理工大学低温生物医学技术研究所，上海200093 [2]上海第二医科大学第九人民医院整形外科，上海市组织工程重点实验室，上海200011 |
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| 摘 要： | 本文采用升降式程序降温仪对人胚胎干细胞进行了程序降温保存，并探讨和比较了降温速率、置核温度、保护剂和投入液氮前温度对冻存复苏后胚胎干细胞的存活率、活力及分化特性的影响。结果表明：采用Me2SO 血清 DMEM(体积比为1：3：6)的保护剂，从0℃开始，以0．5℃／min的速率对细胞悬液降温；至-10℃时对其进行置核，并于-35℃时将其快速投入液氮中保存，复温后效果最佳。冻存复温后细胞存活率可达81．8％，复苏后的胚胎干细胞形态和集落生长方式都与冻前的生长形态相同，且胚胎干细胞标志之一碱性磷酸酶(AKP)反应阳性，同时染色体组型仍正常。
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| 关 键 词： | 人胚胎干细胞 实验研究 保存 程序 细胞存活率 碱性磷酸酶 染色体组型 降温速率 细胞悬液 生长形态 生长方式 细胞形态 保护剂 升降式 核温度 体积比 min 复苏 冻存 液氮 投入 复温 分化 活力 血清 集落 |
Effective cryopreservation of human embryonic stem cells by programmed freezing] |
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| Peng Fei Yang,Hsiao Chien Tsung,Qi Kang Cheng,Tse Chao Hua,Chun Fang Wu,Yi Lin Cao.Effective cryopreservation of human embryonic stem cells by programmed freezing][J].Acta Biologiae Experimentalis Sinica,2005,38(3):247-256. |
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| Authors: | Peng Fei Yang Hsiao Chien Tsung Qi Kang Cheng Tse Chao Hua Chun Fang Wu Yi Lin Cao |
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| Institution: | Institute of Cryomedicine and Cryogenics, Shanghai University of Science and Technology, Shanghai 200093. |
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| Abstract: | Cryopreservation of human embryonic stem cells is an important and unsolved problem. A computer-controlled programmable cooler was used in the preservation of ES cells. Several effects have been experimentally studied, which include the cooling rates, the temperature of seeding, the temperatures before the samples being plunged into liquid nitrogen, and the cryoprotective agents. It was found that the favorable constitution of cryoprotective agents was Me2SO+ FBS+DMEM(1:3:6, v/v/v) with cooling protocol of -0.5 degrees C/min from 0 degrees C to -35 degrees C (seeding at -10 degrees C), and being plunged into the liquid nitrogen immediately. The high survival rate (81.8%) was obtained. |
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