冷冻保存后大鼠胰岛细胞与小肠黏膜下层的共培养

目的探讨猪小肠黏膜下层(SIS)对冷冻保存后大鼠胰岛细胞的支持和保护作用。方法分离纯化后的Wistar大鼠胰岛细胞经冷冻保存1月后分为SIS组和对照组,在RPMI1640培养液培养1周后分别测定两组的胰岛细胞回收率,观察胰岛细胞形态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)%,较游离组(60.6±3.3)%显著提高(P<0.05),胰岛形态较对照组完整。在高糖(16.7mmol/L)刺激下,SIS组胰岛素分泌量较游离组增高[(23.7±1.6)vs(12.5±1.1)mU/L,P<0.05)]。在含有茶碱的高糖溶液中,SIS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材料,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。

Co-culture of cryopreserved rat islets with small intestinal submucosa

OBJECTIVE: To investigate the feasibility and benefits of co-culture of cryopreserved islets with small intestinal submucosa (SIS). METHODS: Purified rat islets cryopreserved for one month were divided into SIS group and control group, and after culture in standard islet culture media RPMI1640 for 1 week, the morphology and function of the islets were assessed. RESULTS: The SIS protects the fragile islets from damage by cryopreservation, and increased the recovery from (60.6-/+3.3)% to (91.7-/+1.8) % (P<0.05). Compared with the control group, incubation of the islets of the SIS group in high-glucose (16.7 mmol/L) solution resulted in significantly enhanced insulin secretion (23.7-/+1.6 vs 12.5-/+1.1 mU/L, P<0.05). When the islets were incubated in high-glucose solution containing theophylline, the calculated stimulation index of SIS group was about 3-fold higher than that of the control group. CONCLUSION: Co-culture of cryopreserved rat islets with SIS can increase the recovery of islet cells and improve their function.