【摘要】 目的通过改进成人胰岛细胞消化分离技术,探索成人胰岛细胞分离纯化的有效方法。方法分别采用改进的手工消化法(n=12);及采用自制的胰岛自动分离装置(n=10)进行成人胰岛分离。比较两组间胰岛分离产量,胰岛素刺激释放试验检测胰岛细胞功能。结果自动消化组每克胰腺胰岛产量(6410.55±1871.6)IEQ/g胰腺,明显高于手工消化法(4428.4±1201.7)IEQ/g胰腺(纯化后)(,P<0.01)。自动消化组高糖刺激后胰岛素释放量为(4.31±0.24)μU/insulin/IE/h,刺激指数为(2.52±0.39)μU/IE/h,均高于手工消化组(4.04±0.23)μU/insulin/IE/h(,1.99±0.53)μU/IE/h(,P<0.05)。结论采用自动消化法分离成人胰岛产量较高,功能良好。胰腺自动分离装置可望进一步推广应用。更多还原

【Abstract】 [Objective] We developed an original automated digestion system and investigated the utility of this system on human pancreatic islet isolation. [Methods] A total of 22 pancreata were divided into two groups: group A (n=12), pancreata were digested using our previous modified manual digestion method, group B (n=10), based on Ricordi′s apparatus we developed an automated system for digestion of the pancreas of human with the cooperation of South China University of Science and Technology, 10 adult cadaveric pancreata were used for islet isolation with our new system. islets were purified on Euro-Ficoll gradient solution. Donor age and recovery of donor pancreas were similar in both groups. [Results] Total islet equivalent number and islet equivalent number per gram pancreas were significantly higher in group B [(283706.2±59664.5) IEQ and (6410.55±1871.6) IEQ/g] than in group A (175381.3±28439.8) IEQ, (4428.4±1201.7) IEQ/g, (P<0.01). Islet purity(84% vs 84%) and viability showed no differences between the two groups. During high glucose stimulation, islets isolated using developed Ricordi′s automated digestion method secreted more insulin than those islets isolated using modified manual digestion method [(4.31±0.24) μU/insulin/IE/hr vs. (4.04±0.23) μU/insulin/IE/hr]. The calculated stimulation index for the group A had a mean of (1.99±0.53), whereas the stimulation index for the group B had a mean of (2.52±0.39) (P <0.05). [Conclusion] The developed Ricordi′s automated digestion method improves islet yield and functional viability of the isolated islets. We recommend this method for human islet isolation.更多还原