改良的胶原酶滑膜细胞分离培养法  

A modified collagenase-digestive method for synovial cell culture

作　　者：彭卫华[1] 侯凡凡[1] 蒋建平[1] 张训[1] 

机构地区：[1]第一军医大学南方医院肾内科,广东广州510515

出　　处：《第一军医大学学报》2000年第5期469-470,共2页Journal of First Military Medical University

基　　金：国家自然科学基金项目(39770349)

摘　　要：目的 探讨一种改良的滑膜细胞培养法。方法 修剪、收集关节滑膜层，依次以胶原酶、0.2%胰蛋白酶、0.2%胰蛋白酶-0.02%EDTANa2消化，100目筛网过滤，D-hanks液洗网后培养，行显微镜、电镜、ABC法免疫组化鉴定。结果 培养的细胞具有成纤维细胞样形态和特征，免疫组化染色示vimentin阳性、CD68阴性，纯度达98%以上，符合B型滑膜细胞特点。结论 改良的胶原酶消化法是一种有效的滑膜细胞培养方法。Objective A modified digestive method was developed to obtain type B synovial cells. Methods Human joint synovial layers were cut into small pieces and digested successively with collagenase, 0.2% trypsin, and 0.2% trypsin-0.02% EDTANa2. Cell suspension was passed through a 100 mesh, washed with D-hanks solution and cultured in DMEM containing 10% fatal bovine serum. Cultured cells were characterized by microscope, electroscope and immunohistochemical analysis. Results The cultured cells showed the shape and feature of fibroblast-like cells, positively stained for vimentin and negatively stained for CD68 (with a purity≥98%), which were confirmed to be human joint type B synovial cells. Pure and sufficient synovial cells were obtained only by one-time digestion. Conclusion The modified collagenase-digestive method is an effective method for synovial cell culture.

关 键 词：细胞培养 滑膜细胞 胶原酶 透析相关性泻粉样变 

分 类 号：R459.5[医药卫生—治疗学]