Effect of Soybean Trypsin Inhibitor on The Isolation and Purification of Rat Islets_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

YANG Yongsheng , ZHANG Xuewen , ZHANG Dan , MENG Zihui , SUN Baozhen , ZHAO Jisheng , WANG Wei

Department of Hepatopancreatobiliary Surgery, China-Japan Union Hospital of Jilin University, Changchun 130033, China;

Corresponding author：

Keywords：

Soybean trypsin inhibitor; Islet isolation; Islet graft

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Objective To study the effect of soybean trypsin inhibitor (STI) on the rat islets’ yield and function during the process of isolation and purification. Methods The rats were divided into experiment group and control group according to whether STI was put into the collagenase. STI (2.0 mg/ml) was put into the collagenase digestive juice of the experiment group and none to the control group. For both of two groups, islets were isolated by situ perfusion collagenase into the rat pancreas and they were purified by un-continuous gradient centrifugation with Ficoll-400. The quantities of the obtained rat islets before and after purification were recorded, and the morphosis and function of the purified rat islets were tested, then their vivo function were observed after islets plantation. Results After digest and before purification, there was no obvious deviation of the obtained islets quantity between two groups 〔(624±38.2) IEQ vs (586±37.7) IEQ, P＞0.05〕; After purification, there were significant deviation in the islets quantity 〔(408±28.3) IEQ vs (189±27.1) IEQ, P＜0.05〕 and purity quotient 〔（93±2.4）% vs （75±2.1）%, P＜0.05〕. For two groups, there was no obvious deviation of the obtained islets in insulin stimulation and secretion experiment as well as their vivo function experiment. Conclusion The ultimate yield and purity quotient of the rat islets can be obviously improved by using collagenase digestive juice with SIT in situ perfusion on the rat pancreas, and it has no obvious effect on the islets function.

Citation： YANG Yongsheng,ZHANG Xuewen,ZHANG Dan,MENG Zihui,SUN Baozhen,ZHAO Jisheng,WANG Wei,.. Effect of Soybean Trypsin Inhibitor on The Isolation and Purification of Rat Islets. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(11): 813-816. doi: Copy

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1. Ricordi C, Lakey JR, Hering BJ. Challenges toward standardization of islet isolation technology[J]. Transplant Proc, 2001; 33(1-2)∶1709.
2. London NJ, Thirdborough SM, Loftus IM， et al. Renal subcapsular islet transplantation in the rat: a comparison of three techniques[J]. Diabetes Res, 1991; 16(2)∶81.
3. Nagata N, Gu Y, Hori H, et al. Evaluation of insulin secretion of isolated rat islets cultured in extracellular matrix[J].Cell Transplant, 2001; 10(4-5)∶447.
4. Avila JG, Wang Y, Barbaro B, et al. Improved outcomes in islet isolation and transplantation by the use of a novel hemoglobin-based O2 carrier[J]. Am J Transplant, 2006; 6(12)∶2861.
5. Xiaohui T, Wujun X, Xiaoming D, et al. Small intestinal submucosa improves islet survival and function in vitro culture[J]. Transplant Proc, 2006; 38(5)∶1552.
6. White SA, Hughes DP, Contractor HH, et al. An investigation into the distribution of different collagen types within adult and juvenile porcine pancreata[J]. J Mol Med, 1999; 77(1)∶79.
7. White SA, Djaballah H, Hughes DP， et al. A preliminary study of the activation of endogenous pancreatic exocrine enzymes during automated porcine islet isolation[J]. Cell Transplant, 1999; 8(3)∶265.
8. Heiser A, Ulrichs K, Müller-Ruchholtz W. Isolation of porcine pancreatic islets: low trypsin activity during the isolation procedure guarantees reproducible high islet yields[J]. J Clin Lab Anal, 1994; 8(6)∶407.
9. Basir I, van der Burg MP, Scheringa M, et al. Improved outcome of pig islet isolation by Pefabloc inhibition of trypsin[J].Transplant Proc, 1997; 29(4)∶1939.
10. Traverso LW, Abu-Zamzam AM. Activation of pancreatic proteolytic enzymes by commercial collagenases[J].Transplantation, 1978; 25(4)∶226.
11. 姚磊，吴德全，李国东，等. 乌司他丁在大鼠胰岛分离过程中的保护作用 [J]. 中华器官移植杂志， 2005; 26（12）∶732.
12. 白日星，宋茂民. 预防性添加胰蛋白酶抑制剂对成年猪胰岛分离的影响 [J]. 中国普外基础与临床杂志, 2005; 12（3）∶275.
13. Lakey JR, Helms LM, Kin T, et al. Serine-protease inhibition during islet isolation increases islet yield from human pancreases with prolonged ischemia[J]. Transplantation, 2001; 72(4)∶565.
14. 袁勤生主编. 现代酶学 [M]. 上海: 华东理工大学出版社， 2001∶107~119.
15. Salvalaggio PR, Deng S, Ariyan CE, et al. Islet filtration: a simple and rapid new purification procedure that avoids ficoll and improves islet mass and function[J].Transplantation, 2002; 74(6)∶877.
16. 张剑, 陈规划, 翁建平, 等. 抗CD40L单克隆抗体在异种胰岛移植排斥反应中的作用 [J]. 中国普外基础与临床杂志, 2008; 15(5)∶329.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of Soybean Trypsin Inhibitor on The Isolation and Purification of Rat Islets

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