【作者】 姜蕾； 胡军祥； 姜玉新；

【Author】 JIANG Lei, HU Jun-xiang, JIANG Yu-xin (The college of life science ,Zhejiang University,Hangzhou 310012,China)

【机构】 浙江大学生命科学学院；

【摘要】 胚胎神经细胞的玻璃化冻存是目前低温生物学研究的重要内容。该文以孕17-19 天的SD大鼠为实验材料,研究了玻璃化的预平衡时间和温度对胚胎神经细胞的存活率、细胞活力、生物发光、H2O2含量等方面的影响。结果表明:玻璃化冻存的预平衡以0℃,分三步平衡30分钟为宜,胚胎神经细胞的存活率可达56．9％。用VSB(20．5％(W／V)DMSO,15． 5％(W／V)乙酰胺,4．5％(W／V)丙二醇,5．5％(W/V)2,3-丁二醇以及6％(W／V)聚乙二醇(MW 6000))作为玻璃化溶液对胚胎神经细胞冻存后,细胞生物发光和H2O2含量随着冻存天数的延长而增加,细胞活力则随着冻存天数的延长而降低,但15天后基本都能达到稳定的状态,且在室温下平衡后的低温损伤更为明显,提示在玻璃化冻存过程中预平衡对胚胎神经细胞的活力具有一定的影响。更多还原

【Abstract】 Cryopreservation of embryonic neurocytes by vitrification is the important content in the cryobiology at present. In this study, pregnant SD rats (17-19days) were used for the research on the changes of the function of embryonic neurocytes after vitrification under the conditions of the different durations and temperatures of pre - equilibration, including the changes of viability rate, vitality of cell, bioluminescence and the content of H2O2. The experiment results showed: the maximum survival rate (56. 9%) was attained by three steps of equilibration for 30 minutes at 0℃.The bioluminescence and the content of H2O2 were increased gradually, whereas the vitality of cell decreased with the prolongation of the cryopreservation, using VSB (20.5%(W/V)Dimethyl sulfoxide , 15. 5%(W/V)Acetamide, 4.5%(W/V)Propane glycol, 5. 5%(W/V)2,3 - butylene glycol and 6%(W/V)Polyethylene glycol (MW 6000)) as vitrification solution. And the stable states all achieved after 15 days of cryopreservation basically. It was more obvious that the damnifications were done at room temperature.It was indicated that the pre - equilibration had the significant effects on the vitality of embryonic neurocytes after the cryopreservation by vitrification to a certain degree.更多还原