成年猪胰岛的分离与纯化

Isolation and purification of islets of langerhans from adult pigs

目的 研究成年猪胰岛的分离与纯化方法。方法 胰岛的分离采用导管内连续灌注胶原酶消化法 ,纯化采用Nycodenz连续密度梯度离心。 结果 消化后每个胰腺平均可获得 (3970 0 0±185 96 3)个胰岛或 (986 0± 3 0 43)个胰岛 /克胰腺组织 ,纯化后为 (1480 0 0± 890 0 0 )个胰岛 /胰腺或(3 75 0± 493)个胰岛 /克胰腺组织 ,纯度 >95 %。体外培养示胰岛对糖刺激反应良好 ;免疫组织化学检查证实胰岛细胞存活。结论 经本法获得的胰岛具有较高的纯度及良好的功能 ,可为临床提供可靠的移植物来源。

Objective To study the method for the isolation and purification of islets from adult pig. Methods Islets were isolated using digestion by ductal injection of collagenase, and purified by centrifugation on Nycodenz gradients. Results From each organ, ( 397?000 ± 185?963 ) islets, corresponding to ( 9?860 ± 3?043 ) islets per gram of pancreas, were obtained. After purification the recovery was ( 148?000 ± 89?000 ) islets/pancreas ( 3?750 ±493 )islets per gram of pancreas; the purity of the final preparation was >95?%. The culture of the islets in vitro confirmed the viable cells. Conclusions The islets produced by this method could provide a possible source of purified, physically functional graft for clinical transplantation.