超声波植冰对L-02肝细胞低温保存的影响  

Effect of Ultrasonic Ice Seeding on Cryopreservation of L-02 Hepatocyte

作　　者：李维杰[1] 宋立勇 刘宝林[1] 宗果 Li Weijie;Song Liyong;Liu Baolin;Zong Guo(Institute of Biothermal Science and Technology,University of Shanghai for Science and Technology,Shanghai,200093,China;Shanghai Horizon Medical Co.,Ltd.,Shanghai,200120,China)

机构地区：[1]上海理工大学生物系统热科学研究所,上海200093 [2]上海明悦医疗科技有限公司,上海200120

出　　处：《制冷学报》2021年第4期158-166,共9页Journal of Refrigeration

基　　金：国家自然科学基金(51776130)资助项目;上海市上海理工大学医工交叉项目(1020308412);上海市科委战略新兴产业引导专项(18441909700)资助

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Me 2SO对肝细胞存活率的影响。结果表明:当体积分数为0%~10%的Me 2SO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0.0329~0.4316 W/cm^(2)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0.4316 W/cm^(2)时,肝细胞存活率显著下降;使用体积分数为5%的Me 2SO在-7℃进行超声波植冰,超声波强度为0.1041 W/cm^(2)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Me 2SO用量。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 Me 2SO on the survival rate of hepatocytes were explored.The results show that when Me 2SO 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 0.0329-0.4316 W/cm^(2),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 0.4316 W/cm^(2),the hepatocyte survival rate decreased significantly;ultrasonic ice seeding with Me 2SO at a volume concentration of 5%at-7℃and an ultrasonic intensity of 0.1041 W/cm^(2)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 Me 2SO in the solution.

关 键 词：肝细胞 过冷度 低温保存 超声植冰 

分 类 号：TB61.1[一般工业技术—制冷工程] R318.52[医药卫生—生物医学工程]