微波复温对玻璃化冻存大鼠胚脑细胞活性的影响

Effects of microwave thawing on viability of rat embryonic cerebral cells cryopreserved by vitrification

以孕17～19d的SD大鼠为材料,对胚脑细胞进行玻璃化深低温保存,在冻存3、10、20、30、60d后,胚脑细胞样品分别采用常规的38℃水浴复温和微波复温即在300mL10℃水中,控制微波功率为800W,频率2450MHz,复温时间80s.检测胚脑细胞的功能变化,比较两种复温方法对细胞活性的影响.结果表明:胚脑细胞玻璃化冻存3～30d,细胞的回收率、存活率、细胞内琥珀酸脱氢酶(SDH)、超氧化物歧化酶(SOD)活性都有下降的趋势,过氧化氢(H2O2)含量显著增加,但冻存30d后这些指标保持基本稳定,与第60d比较无显著差异(P>0.05).微波复温组的细胞存活率,细胞内SDH、SOD的活性高于水浴复温组,细胞内H2O2含量明显低于水浴复温组.说明微波复温能够提高复温速率,对胚脑细胞的损伤要低于水浴复温.

The embryonic cerebral cells (ECC) of pregnant SD rats (17-19 days) were cryopreserved by vitrification.After cryopreservation for 3,10,20,30 and 60 days, the individual ECC samples were separately thawed in 38 ℃ water bath or by microwave treatment in 300 mL 10 ℃ water under microwave power of 800W, frequency of 2450MHz and thawing duration of 80s, following which the viability of ECC was assayed.The experimental results showed that during cryopreservation of 3-30 days, the recovery rate and the survival rate of the cells,and the activity of succinate dehydrogenase(SDH)and superoxide dismutase (SOD) of ECC were significantly depressed, whereas the hydrogen peroxide(H_2O_2)content of ECC was markedly increased.After cryopreservation for 30 days these indexes remained relatively stable so that no significant difference (P>0.05) was found between in these indexes before and after being preserved for 60 days. The cell survival rate, the activity of SDH and SOD of ECC subjected to microwave thawing were higher than those of ECC subjected to water bath thawing, whereas the H_2O_2 content of ECC of the former was lower than that of the latter. It was indicated that damage to the microwave-thawed ECC was lighter than damage to water bath-thawed ECC.