【摘要】 【目的】采用不同的分离或消化方法 ,探讨如何提高大鼠胰岛细胞分离纯化后的收获量和质量。【方法】将 2 5只供体SD雄性大鼠依胰腺分离或消化的方法不同随机地分为 5组 :①A组 :胆总管灌注胶原酶P ;②B组 :胆总管灌注Hanks液 ;③C组 :胆总管不灌注液体 ;A、B、C 3组胰腺直接分次消化 ;④D组 :胆总管不灌注 ,胰腺剪碎后分次消化 ;⑤对照组 :胆总管灌注胶原酶P ,胰腺直接单次消化。将分离的胰岛沉淀物用Ficoll 4 0 0纯化并培养 ,再把胰岛细胞移植于糖尿病裸鼠受体腹腔内。【结果】纯化后的胰岛细胞在收获量上A和B组明显多于对照组或C组 ,而D组最低 ;A、B和D 3组的纯度均高于对照组或C组 ;在胰岛成活率方面 ,A、B、C 3组均高于对照组或D组。移植于糖尿病裸鼠受体内的大鼠胰岛细胞均可逆转高血糖状态。【结论】经胆总管灌注胶原酶后的大鼠胰腺用分次消化的方法可提高胰岛细胞的收获量、纯度和活性。更多还原

【Abstract】 Objective To investigate how to improve the yield and quality of rat islets in isolation and purification. Methods 25 male SD rats (islet donors) were randomly divided into 5 groups based on different methods of separation or collagenase digestive approach. ①Islets were isolated using two times digestion by pancreatic ductal injection of collagenase P (group A). ②Hanks solution was injected through the common bile duct into the pancreas, and the distended pancreas directly digested by collagenase P for two times (group B). ③There was no solution to be injected through the common bile duct into the pancreas, and the pancreas digested directly by collagenase P for two times (group C). ④The common bile duct didn’t need to be injected. The pancreas was directly cut into small pieces, and digested by collagenase P for two times (group D). ⑤The pancreas was directly digested by collagenase P for one time after pancreatic ductal injection of collagenase P (control group). Islets were purified by Ficoll-400 density gradient and cultured, and transplanted into the abdominal cavity of diabetic nude mice. Results The yield of purified islets was much more in group A or B than in group C or control group, and that was the least in group D. The purity of group A, B, and D was significantly higher than that of control group or group C, respectively. The survival rate of islets in group A, B, and C was significantly higher than that of control group or group D, respectively. Transplanted rat islets could reverse hyperglycemia in the diabetic nude mice. Conclusion After injection of collagenase, the distended pancreas is removed and digested by collagenase for two times. This method can improve the yield, purification and viability of purified rat islets.更多还原