Isolation of the elusive pig islet.
Isolation of the elusive pig islet.
复制标题
难以捉摸的猪胰岛的分离。
作者：
C. Ricordi;C. Socci;A. Davalli;C. Staudacher;P. Baro;A. Vertova;I. Sassi;F. Gavazzi;G. Pozza;V. Carlo
本研究描述了一种大规模分离猪胰岛的方法。报告的结果来自15个连续的分离株。从没有热缺血的活体动物身上取出腺体，用改进的人类胰岛自动分离方法消化胰腺。平均每个胰腺可分离690,000个+/-279,429个胰岛，相当于每克胰腺10,360+/-4034个胰岛，体积为714+/-480mm3。纯化后回收率为255,000+/-32,407个胰岛，相当于每克胰腺4,000+/-567个胰岛。最终制备的胰岛纯度为80%~95%。胰岛素含量纯化前平均为146.8+/-78U，纯化后平均为71+/-53U。将10mm3的最终制剂移植到7只患糖尿病的裸鼠的肾包膜下后，其中6只小鼠出现了正常血糖。移植后30天，所有病例的移植肾切除后均迅速恢复到糖尿病状态。这种方法可以提供大量完整的猪胰岛，为通过免疫改变和免疫隔离程序预防猪胰岛移植排斥反应的初步研究提供了可能。
This study describes a procedure for large scale isolation of swine islets. The reported results are from 15 consecutive isolations. The glands were removed from live animals with no warm ischemia, and the pancreata were digested by a modification of the automated method for human islet isolation. It was possible to separate an average of 690,000 +/- 279,429 islets per pancreas corresponding to 10,360 +/- 4034 islets per gram of pancreas with a volume of 714 +/- 480 mm3. After purification the recovery was 255,000 +/- 32,407 islets corresponding to 4,000 +/- 567 islets per gram of pancreas. Purity of the final preparation was 80% to 95% islets. Insulin content resulted in an average of 146.8 +/- 78 U before purification and 71 +/- 53 U after purification. After a 10 mm3 aliquot of the final preparation was transplanted under the renal subcapsular space of seven nude mice with diabetes, normoglycemia occurred in six of the mice. Thirty days after transplantation, nephrectomy of the kidneys bearing the grafts produced a rapid return to the diabetic state in all cases. This method makes it possible to provide large numbers of intact swine islets for preliminary studies of prevention of the rejection of pig islet xenograft by immunoalteration and immunoisolation procedures.