人翼状胬肉细胞的分离和培养

Primary Culture of Human Pterygia Cells in Vitro

目的:探索人翼状胬肉细胞的体外分离和培养方法。方法:取原发翼状胬肉50例,无菌条件下将胬肉组织剪成1×11／mm2的组织块,37℃环境中 0．025％Ⅱ型胶原酶消化20 min,0．05％胰蛋白酶消化10 min,1500 rpm离心10 min,将收集到的细胞用内皮细胞条件培养基制成悬液后进行接种培养,经机械刮除和密度梯度离心法进行纯化,采用CD31、CD34、Ⅷ因子免疫组化和透射电镜方法进行鉴定,设阴性和空白对照。结果:经原代培养后可得到人翼状胬肉成纤维细胞和内皮细胞样细胞,胞浆内分别含有VIM 和CD31、CD34及W-P小体。结论:本实验培养获得了人翼状胬肉成纤维细胞和内皮细胞样细胞,方法简便,可操作性强, 为进一步研究翼状胬肉的发病机制奠定了坚实的基础。

Purpose：To probe a selective method of isolate and culture of human pterygia cells in vitro. Methods：Fifty primary pterygia were got by operation and cut to clips less than 1 ×1mm^2, segments were digested by collagenase Ⅱ for 20 minutes and by trypsin for 10 minutes under 37℃, and then centrifugated for 15 minutes by 1 500 rpm. The collected cells were cultured in conditioned medium for endothelial cells. After purified by scrape and density gradient centrifugation, the ceils were detected and identified by immunohistochemistry and electron microscope. Results：The purity of cultured fibroblasts and endothelial cell type cells were more than 90%. CD31, CD34, and Weibel-Palade bodies were performed in these cells cytoplasm. Conclusion：We obtained the method of culturing human pterygia fibroblasts and endothelial cell type cells and it is a feasible and convenient approach.