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胶原酶金加BP蛋白酶用于从人胰腺中分离胰岛的评估。

Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata.

作者信息

Khiatah Bashar, Tucker Amber, Chen Kuan-Tsen, Perez Rachel, Bilbao Shiela, Valiente Luis, Medrano Leonard, Rawson Jeffrey, Forouhar Elena, Omori Keiko, Kandeel Fouad, Qi Meirigeng, Al-Abdullah Ismail H

机构信息

a Department of Translational Research and Cellular Therapeutics , Diabetes and Metabolism Research Institute, Beckman Research Institute of the City of Hope , Duarte , CA , USA.

出版信息

Islets. 2018 Mar 4;10(2):51-59. doi: 10.1080/19382014.2017.1417716. Epub 2018 Feb 2.

Abstract

Selection of enzymes for optimal pancreas digestion is essential for successful human islet isolations. The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15). The isolation outcomes were assessed by islet counting, viability, glucose-stimulated oxygen consumption rate (OCR), and successful graft-rate following transplantation in diabetic NOD scid mice. The pancreas donor characteristics were not significantly different between the tested enzyme groups regarding their BMI, pancreas weight, cold ischemia time (CIT) and HbA1c. The results show that digested tissue volume was not statistically significant between the VitaCyte enzyme (34.25 ± 5.4 mL) and the Roche enzyme (55.25 ± 3.42 mL, p = 0.073), however, this was significant with Serva enzyme (64.07 ± 7.95 mL, p = 0.020). Interestingly, the islet yields were not statistically different between all enzyme groups. Moreover, when islets were transplanted into NOD scid mice, the reversal rate of diabetes for the VitaCyte enzyme group was similar to all enzyme groups. In conclusion, the effectiveness of Collagenase Gold plus BP protease is comparable to the MTF C/T and the Collagenase NB1/NP enzymes; the low cost could facilitate the use of more pancreata for islet isolations.

摘要

选择用于胰腺最佳消化的酶对于成功进行人胰岛分离至关重要。本研究的目的是通过将胶原蛋白酶 Gold 加 BP 蛋白酶(VitaCyte)(n = 8)与两种市售酶 Liberase MTF C/T(罗氏)(n = 48)和胶原蛋白酶 NB1/NP(塞尔瓦)(n = 15)进行比较,来评估其功效和结果。通过胰岛计数、活力、葡萄糖刺激的氧消耗率(OCR)以及在糖尿病 NOD 无胸腺小鼠移植后的成功移植率来评估分离结果。在测试的酶组之间,胰腺供体的特征在体重指数、胰腺重量、冷缺血时间(CIT)和糖化血红蛋白方面没有显著差异。结果表明,VitaCyte 酶(34.25 ± 5.4 mL)和罗氏酶(55.25 ± 3.42 mL,p = 0.073)之间消化组织体积无统计学显著差异,然而,与塞尔瓦酶(64.07 ± 7.95 mL,p = 0.020)相比有显著差异。有趣的是,所有酶组之间的胰岛产量无统计学差异。此外,当将胰岛移植到 NOD 无胸腺小鼠中时,VitaCyte 酶组的糖尿病逆转率与所有酶组相似。总之,胶原蛋白酶 Gold 加 BP 蛋白酶的有效性与 MTF C/T 和胶原蛋白酶 NB1/NP 酶相当;低成本可能有助于使用更多胰腺进行胰岛分离。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6150/5895173/5d603d228be3/kisl-10-02-1417716-g001.jpg

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