Abstract: To study the method of islet isolation and purification in adult rats and make preparation for clinical use, collagenase was injected into pancreatic duct followed by incubation in water bath to digest the pancreas and isolate islet. Then discontinuous gravity gradient purification was used to purify the islet. Secretion of insulin was assessed after stimulation of glucose in tube. 938±218 islets were procured after isolation and they were intact in morphology. 802±181 is lets were recovered after purification. The average purity was (68±11)%, recovery rate was (85.9±5.9)%. The islets were morphologically intact after the purification. When inspected by glucose stimulation, the secreted insulin concentration was (2.53±0.18) mIU/L when stimulated by low concentration glucose and (5.41±0.31) mIU/L by high concentration glucose. The difference was very significant(P=0.001). C-pep was (7.51±1.09) ng/L when stimulated by low concentration glucose and (8.56±1.06) ng/L by high concentration glucose. The difference was very significant (P=0.006). Conclusion: More islets (802±181) can be procured with good function and shape by using the method of injecting collagenase into pancreatic duct followed by incubation in water bath and purification using discontinuous gravity gradient. This sets the foundation of preparation for clinical transplantation.

Key words: islet, isolation, purification