摘要
目的:探讨猪小肠黏膜下层(SIS)对冷冻保存后的胰岛细胞的支持和保护作用.方法:将分离纯化后的W istar大鼠胰岛细胞经冷冻保存1 mo后分为SIS组和对照组,在RPMI 1640培养液培养1 wk后分别观察胰岛细胞形态,测定两组的胰岛细胞回收率并进行胰岛素刺激释放试验.结果:SIS组胰岛细胞回收率为(91.7±1.8)%,较对照组(60.6±3.3)%显著提高(P<0.05),胰岛形态较对照组完整.在高糖(16.7 mmol/L)刺激下,SIS组胰岛素分泌量较对照组增高[(23.7±1.6)mU/Lvs(12.5±1.1)mU/L,P<0.05].在含有茶碱的高糖溶液中,微囊组的刺激指数为对照组的3倍.结论:SIS作为一种天然的细胞外基质,可显著提高胰岛细胞冷冻保存后的回收率,减少冻存胰岛细胞复苏时的死亡并改善胰岛细胞的功能.
AIM： To investigate the protective effect of small intestinal submucosa （SIS） on rat islet cells after cryopreservation. METHODS： The purified rat islet cells were cryopreserved for 1 month. After cryopreservation, the cells were separated into 2 groups： SIS group and control group. The 2 groups were cultured in standard islet cell culture media of RPMI 1640 for 1 week..M＇ter that, the islet cells of the 2 groups were stained with dithizone and counted. Recovery of islet cells was determined and viability of islet cells was assessed by the glucose challenge test. RESULTS： The SIS protected the fragile islet cells from freezing damage, which was demonstrated through the significant difference in the recovery rate between the SIS group （91. 7 ± 1.8）% and the control group （60.6 ± 3.3 ） % （ P 〈 0.05 ）. In the situation of the incubation with high glucose solution （ 16.7 mmol/L） , the insulin secretion in the SIS group showed significantly increased as compared with the control group [ （23.7 ± 1.6） mU/Lvs （12,5±1.1） mU/L, P〈0.05], When the islet cells were placed in the high glucose solution containing theophylline. the calculated stimulation index of the SIS group was about 3 times as high as that of the control group. CONCLUSION： Coculture of rat islet ceils after cryopreservation with SIS is a promising way to increase recovery and improve function of islet cells in the process of thaw.
出处
《第四军医大学学报》
CAS
北大核心
2006年第15期1373-1375,共3页
Journal of the Fourth Military Medical University
基金
教育部科学技术重点项目(104169
23004-2007)
关键词
冷冻保存
回收率
胰岛功能
小肠黏膜下层
cryopreservation
recovery rate
islet function
small intestinal submucosa
