【Abstract】 Objective To explore good methods for isolation and purification of rat islets. Methods The islets
were isolated from male SD rat pancreata by a collagenase perfusion method and purified by a modified method: added 4 kinds of Euro-Ficoll of different densities (F1: D=1.132, F2: D=1.108, F4: D=1.069, F5: D=1.023), discontinuous density gradient centrifuge the tube at 2 000 r/min for 20 minutes at 4℃ , then the islets between F1 and F2 were collected. The purity of islets was assessed by dithizone staining with islets counted and scored for size. Islets viabil ity was assessed by fluorescin diacetate / propidium iodide. The function of purified islets was judged by the test of insul in release and islets transplantation. Results After an improved method for optimized isolation and purification, (920±122) IEQ purified islets were obtained from one rat. Both the purity and viabil ity of islets were over 90%. The amount of insul in secretion was (18.25±0.32) mU/L and (36.70±3.57) mU/Lat 2.2 mmol/ L and 22.2 mmol/L concentration of glucose respectively, there was significant difference between the two phases
(P lt; 0.05). The insul in release index was 2.01±0.15. Under 1 000 IEQ islets transplantation, the normal glucose level could beremained in diabetic rats. Conclusion High purity and high viabil ity islet cells can be got through improved collagenase perfusion and centrifugation on gradients method.
Citation： YUAN Yu,CONG Cong,ZHANG Jing,WEI Lingl ing,LI Shengfu,JIN Xi,MAI Gang,LI Youping,CHENG Jingqiu,LU Yanrong. IMPROVED METHOD FOR OPTIMIZED ISOLATION AND PURIFICATION OF RAT ISLETS AND IDENTIFICATION OF FUNCTION. Chinese Journal of Reparative and Reconstructive Surgery, 2008, 22(1): 75-79. doi: Copy