大鼠胰岛细胞原代培养方法比较研究

目的：比较减碎消化法和原位灌注消化法在大鼠胰岛细胞原代培养中的优缺点,探讨一种简单易行的大鼠胰岛细胞培养方法。方法：雄性SD大鼠30只随机分成减碎组（CC组,n=15）,胰腺剪为小组织块后,Ⅴ型胶原酶逐级消化分离;原位灌注组（YY组,n=15）,采用胆总管逆行灌注Ⅴ型胶原酶分离胰岛。双硫腙（DTZ）染色检测胰岛纯度,葡萄糖刺激胰岛素释放试验检测胰岛功能。结果：2组获得的胰岛对于葡萄糖刺激均具有良好的分泌活性（P〈0.01）;YY组比CC组获得的胰岛数量多（P〈0.01）,形态较好,等量胰岛的胰岛素释放量亦多（P〈0.05）。结论：胆总管原位灌注Ⅴ型胶原酶消化可以获取数量多,活性和纯度较好的胰岛。

A comparative study on the primary culture of rat islet cells

Objective： To compare the cut-into-pieces digestion method and in situ injection of collagenase V method for isolating rat islet,and to determine a simple and convenient method for primary culture of rat islet cells. Methods： Thirty male SD rats were randomly divided into two groups. In group CC （ n = 15） ,the pancreas were cut into pieces and digested gradually by collagenase V. In group YY （ n = 15） ,collagenase V were injected through bile duct and islets were isolated. The purity of islets was examined by dithizone staining,and their function was evaluated by glucose-stimulated insulin release test. Results： The insulin secretion was both active （ P 0. 01） . Compared with group CC,more islets with better morphology （ P 0. 01） and active release of insulin were obtained in group YY（ P 0. 05） . Conclusion： The method of in situ injection of collagenase V through bile duct can obtain more islets with higher activity and purity.