A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products.
A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products.
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DOI：
10.1097/tp.0b013e318247281b
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发表时间：
2012-04-15
期刊：
影响因子：
6.2
通讯作者：
Hering BJ
中科院分区：
文献类型：
--
作者：
Balamurugan AN;Loganathan G;Bellin MD;Wilhelm JJ;Harmon J;Anazawa T;Soltani SM;Radosevich DM;Yuasa T;Tiwari M;Papas KK;McCarthy R;Sutherland DE;Hering BJ
最佳的酶混合物，最大限度地提高人类胰岛移植产量仍有待确定。在这项研究中，我们评估了8种不同的酶组合（EC），试图提高胰岛产量。EC由来自溶组织梭菌（Ch）的纯化的、完整的或截短的1类（C1）和2类（C2）胶原酶以及来自嗜热解蛋白芽孢杆菌rokko（嗜热菌蛋白酶）或Ch（ChNP）的中性蛋白酶（NP）组成。我们报告了249例人胰岛分离的结果，包括99例死亡供体（研究n=57，临床n=42）和150例慢性胰腺炎胰腺。我们制备了一种新的酶混合物（NEM），由完整的C1和C2胶原酶和ChNP组成，而不是使用嗜热菌蛋白酶。NEM首先在分裂胰腺（n=5）实验中进行测试，然后用于胰岛自体（n=21）和同种异体移植（n=10）。使用多变量分析对8种不同EC的胰岛分离结果进行统计学比较。与其他标准EC相比，NEM始终从胰腺炎（p<0.003）和死亡供体胰腺（p<0.001）获得更高的胰岛产量。使用NEM，来自10个连续胰腺中的8个的胰岛产物满足移植的释放标准，平均6510±2150 IEQ/g胰腺和694，681 ± 147，356总IEQ/移植。在自体分离中，无论纤维化的严重程度如何，NEM从21个胰腺中的19个获得> 200，000 IEQ（平均422，893 ± 181，329总IEQ和5979±1469 IEQ/kg受体体重）。由溶组织梭菌中性蛋白酶和Cizyme高完整C1胶原酶组成的新酶混合物从死亡和胰腺炎胰腺中恢复更高的胰岛产量，同时保留胰岛质量和功能。
The optimal enzyme blend which maximizes human islets yield for transplantation remains to be determined. In this study, we evaluated 8 different enzyme combinations (ECs) in an attempt to improve islet yield. The ECs consisted of purified, intact, or truncated class 1 (C1) and class 2 (C2) collagenases from Clostridium histolyticum (Ch) as well as neutral protease (NP) from Bacillus thermoproteolyticus rokko (thermolysin) or Ch (ChNP). We report the results of 249 human islet isolations, including 99 deceased donors (research n=57, clinical n=42) and 150 chronic pancreatitis pancreases. We prepared a new enzyme mixture (NEM) composed of intact C1 and C2 collagenases and ChNP instead of using thermolysin. The NEM was first tested in split pancreas (n=5) experiments and then used for islet autologous (n=21) and allogeneic transplantation (n=10). Islet isolation outcomes from 8 different Ecs were statistically compared using multivariate analysis. The NEM consistently achieved higher islet yields from pancreatitis (p<0.003) and deceased donor pancreases (p<0.001) than other standard ECs. Using the NEM, islet products met release criteria for transplantation from 8 of 10 consecutive pancreases, averaging 6510±2150 IEQ/g pancreas and 694,681±147,356 total IEQ/transplantation. In autologous isolation, the NEM yielded >200,000 IEQ from 19 of 21 pancreases (averaging 422,893±181,329 total IEQ and 5979±1469 IEQ/kg recipient body weight) regardless of the severity of fibrosis. A new enzyme mixture composed of Clostridium histolyticum neutral protease with CIzyme high intact C1 collagenase recovers higher islet yield from deceased and pancreatitis pancreases while retaining islet quality and function.