微波复温对玻璃化冻存小鼠骨髓有核细胞活性的影响

Effect of Microwave Thawing on Viability of Mice Bone Marrow Cells Cryopreserved by Vitrification

对ICR小鼠骨髓有核细胞进行玻璃化深低温保存,在冻存1、5、10、20、30d后,骨髓有核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。检测骨髓有核细胞的功能变化,比较两种复温方法对细胞活性的影响。结果表明:骨髓有核细胞玻璃化冻存1～20d,其存活率、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20d后这些指标保持基本稳定,与第30d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,对骨髓有核细胞的损伤要低于水浴复温。

The ICR mice bone marrow nucleated cells （BMNC） were cryopreserved by vitrification for 1, 5, 10, 20 and 30 days. Then the individual samples thawed in 38℃ water bath and with microwave treatment, respectively. The viability of BMNC was assayed to compare the influence of the two thawing methods on the viability. The experimental results show that during 1-20 cryopreservation days, the survival rate of the cells, and the activity of succinate dehydrogenase （SDH） and superoxide dismutase （SOD） of BMNC decreased significantly, whereas the hydrogen peroxide （H2O2） content of BMNC increased greatly. After cryopreservation for 20 days these indices remained relatively stable and no significant difference （P〉0.05） was found in these indices before and after being preserved for 30 days. The cell survival rate, the activity of SDH and SOD of BMNC with microwave thawing were higher than those with water bath thawing, whereas the H2O2 content of BMNC with microwave thawing was lower than that with water bath thawing. These findings confirm that microwave thawing may raise the thawing rate and the damage to BMNC by microwave thawing may be samaller than that by water bath thawing.