猫胰岛细胞的分离纯化和培养

目的建立猫胰岛细胞分离纯化和体外培养的方法。方法在自制玻棒持续振荡下常规分离胰岛细胞，进行单层细胞培养和组织块培养。采用差速贴壁法。碘乙酸法、消化时间差法去除培养中的成纤维细胞。动态收集单层细胞培养液， 分别用放射免疫法和比色法检测胰岛素和淀粉酶含量。光镜下观察５～３０ｄ细胞生长情况。分别对培养第５、１５、２１天的 细胞进行葡萄糖负荷试验、组织块培养的细胞进行透射电镜检查。结果猫胰岛细胞在体外培养可保持良好的生物学活 性３周或３周以上（组织培养）。结论　在国内首次建立的猫体外胰岛细胞培养方法可行可靠，体外培养第７～２５ｄ的细 胞形态和功能良好，是进行实验研究的极好材料。

A method for isolating and culturing in vitro feline islet cells

Objective: To establish a method for isolating and culturing feline islet cells. Methods The pancreata taken from l-month-year-old cats were digested with collagenase for mololayer cell and tissue culture respectively, and the fibroblasts were eliminated in the process of cell culture. The content of insulin and amylase in the monolayer cell culture media was measured every 2 d. Morphological observation of the islets cultured in vitro for 5 to 30 d was performed, and the cells undergoing the 2 different cell culture methods were subjected to glucose load test on days 5, 15 and 21 of culture. Results The feline islet cells cultured in vitro could well preserve their biological properties for more than 3 weeks. Conclusion The culture method of the feline islet cells is feasible and reliable and the cultured cells are capable of good performance in subsequent studies.