Vargas F, Vives-Pi M, Somoza N, Alcalde L, Armengol P, Martí M, Serradell L, Costa M, Fernandez-Llamazares J, Sanmartí A, Pujol-Borrell R
Surgery Department, Hospital Universitari "Germans Trias i Pujol," Badalona, Spain.
Transplantation. 1996 Jun 15;61(11):1562-6. doi: 10.1097/00007890-199606150-00002.
Human islet transplantation has a high rate of failure, often due to primary nonfunction, which suggests that islets are damaged during the processing of the pancreas. The preparation of human islets for transplantation is still a complex process that requires large teams of surgical and laboratory personnel. To overcome this problem, we have adopted the use of the IBM 2991 COBE cell separator and a metrizamide/Ficoll density medium that is easy to prepare. Twenty-seven pancreatic glands have been processed using the COBE cell separator, 23 of which were purified in metrizamide/Ficoll gradients and 4 in bovine serum albumin gradients. The results show an improvement of recovery and viability in these preparations when compared retrospectively with manual gradients. More importantly, the time required for purification was shortened to one fourth the usual time and total processing time is about half as long. Moreover, a team of two laboratory staff was regularly able to prepare islets for transplantation, reducing the separation time from 7 hr to 3.5 hr. We conclude that the automatic cell separator and metrizamide-based separation medium are useful modifications of current islet purification methods.
人类胰岛移植的失败率很高，这通常是由于原发性无功能，这表明胰岛在胰腺处理过程中受到了损伤。用于移植的人类胰岛的制备仍然是一个复杂的过程，需要大量的外科和实验室人员团队。为了克服这个问题，我们采用了IBM 2991 COBE细胞分离器和一种易于制备的甲泛葡胺/聚蔗糖密度介质。已经使用COBE细胞分离器处理了27个胰腺，其中23个在甲泛葡胺/聚蔗糖梯度中纯化，4个在牛血清白蛋白梯度中纯化。与传统的手工梯度法相比，这些制备物的回收率和活力有了改善。更重要的是，纯化所需的时间缩短到了通常时间的四分之一，总处理时间约为原来的一半。此外，两名实验室工作人员组成的团队能够定期为移植制备胰岛，将分离时间从7小时缩短到3.5小时。我们得出结论，自动细胞分离器和基于甲泛葡胺的分离介质是对当前胰岛纯化方法的有益改进。
