Bucher Pascal, Mathe Zoltan, Morel Philippe, Bosco Domenico, Andres Axel, Kurfuest Manfred, Friedrich Olaf, Raemsch-Guenther Nicole, Buhler Leo H, Berney Thierry
Cell Isolation and Transplantation Center, Department of Surgery, Geneva University Hospital, Geneva, Switzerland.
Transplantation. 2005 Jan 15;79(1):91-7. doi: 10.1097/01.tp.0000147344.73915.c8.
Biologic characteristics of enzyme products used for islet isolation are critical for the success of islet transplantation. In particular, lot-to-lot variability significantly affects the yields of the isolation procedure. In this study, we have evaluated a new enzyme preparation in which highly purified collagenase can be blended with separately supplied neutral protease in a predetermined ratio.
We compared the results of human-islet isolations performed either with Collagenase NB1 supplemented with Neutral Protease NB (group I, n=9) or with Liberase (group II, n=9).
Endotoxin contents of the enzyme preparations were lower in group I. Total islet yields were similar in both groups, but islet equivalents per gram of pancreas was higher in group I (4,020+/-1,240 vs. 2,360+/-1,350; P<0.05). Islet morphology was improved in group I with significantly higher proportion of free and intact islets (71+/-9% vs. 52+/-14%; P<0.01). In vitro function was improved and apoptosis rate was lower in group I.
This new enzyme blend was as efficient as Liberase in terms of islet yields and showed improvements in islet morphology, viability, and in vitro function. The possibility to control lot-to-lot variability and the low endotoxin contents make Collagenase NB1 a promising product for human-islet isolation.
用于胰岛分离的酶产品的生物学特性对胰岛移植的成功至关重要。特别是，批次间的变异性显著影响分离过程的产量。在本研究中，我们评估了一种新的酶制剂，其中高度纯化的胶原酶可以与单独供应的中性蛋白酶按预定比例混合。
我们比较了使用补充有中性蛋白酶NB的胶原酶NB1（第一组，n = 9）或 Liberase（第二组，n = 9）进行人胰岛分离的结果。
第一组酶制剂的内毒素含量较低。两组的总胰岛产量相似，但第一组每克胰腺的胰岛当量更高（4,020±1,240对2,360±1,350；P<0.05）。第一组的胰岛形态得到改善，游离和完整胰岛的比例显著更高（71±9%对52±14%；P<0.01）。第一组的体外功能得到改善，凋亡率更低。
这种新的酶混合物在胰岛产量方面与 Liberase 一样有效，并且在胰岛形态、活力和体外功能方面有所改善。能够控制批次间变异性以及低内毒素含量使胶原酶NB1成为用于人胰岛分离的有前景的产品。
