| 摘要: |
| 细胞溶液在慢速冷冻过程中会出现过冷现象，较大的过冷度会形成不可控的胞内冰。为减少过冷度，提高细胞存活率，本文提出一种用于L-02肝细胞植冰保存过程的超声波植冰系统，实验研究不同超声波强度、不同预冷温度和不同体积分数Me2SO对肝细胞存活率的影响。结果表明：当体积分数为0%~10%的Me2SO作为保护剂时，使用超声波植冰操作均能显著提高肝细胞冷冻存活率。当超声波强度为0.0329~0.431 6 W/cm2时，肝细胞存活率均在90%以上，组间无显著性差异(p<0.05)，超声波强度大于0.431 6 W/cm2时，肝细胞存活率显著下降；使用体积分数为5%的Me2SO在﹣7 ℃进行超声波植冰，超声波强度为0.104 1 W/cm2时，肝细胞存活率最高，达到(97±2.6)%，与新鲜组无显著性差异；超声波植冰能够显著提高肝细胞存活率，降低保护液中Me2SO用量。 |
| 关键词: 肝细胞 过冷度 低温保存 超声植冰 |
| DOI： |
| 投稿时间：2020-09-25 修订日期：2021-02-08
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| 基金项目:国家自然科学基金(51776130)资助项目。 |
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| Effect of Ultrasonic Ice Seeding on Cryopreservation of L-02 Hepatocyte |
| Li Weijie,Song Liyong,Liu Baolin,Zong Guo |
| (Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology;Shanghai Horizon Medical Co., Ltd.) |
| Abstract: |
| During the slow freezing process, the cell solution is super-cooled, which may cause uncontrollable intracellular ice formation. Ultrasonic ice seeding can reduce super-cooling and improve the cell survival rate. An ultrasonic ice seeding device was designed for the ice seeding preservation of L-02 hepatocytes, and the influence of different ultrasonic intensities, different pre-cooling temperatures, and different volume concentrations of Me2SO on the survival rate of hepatocytes were explored. The results show that when Me2SO with a volume concentration of 0%–10% is used as a cryoprotectant, the use of ultrasonic ice seeding operation can significantly improve the survival rate of hepatocytes during freezing. When the ultrasound intensities are 0.032 9 W/cm2~0.431 6 W/cm2, the hepatocyte survival rate is above 90%, and there is no significant difference between the groups (p<0.05). When the ultrasound intensity is greater than 0.431 6 W/cm2, the hepatocyte survival rate decreased significantly; ultrasonic ice seeding with Me2SO at a volume concentration of 5% at ﹣7 ℃ and an ultrasonic intensity of 0.1041 W/cm2 results in the highest survival rate of hepatocytes, which is (97±2.6)% and is not significantly different from the fresh group. Therefore, ultrasonic ice seeding can significantly improve the survival rate of hepatocytes and reduce the amount of Me2SO in the solution. |
| Key words: hepatocyte super-cooling cryopreservation ultrasonic ice seeding |
