为细胞培养提供单纯结膜上皮的新方法——翻转-固定分离法的实验研究

Ohtaining pure conjunctival epithelium with method of overturn-fixing.

目的 探索一种新的分离结膜上皮与固有层的方法,为结膜上皮细胞的培养提供单纯的结膜上皮组织。方法采用翻转-固定分离法分离结膜上皮与固有层。分离下来的结膜上皮进行组织切片HE染色,以观察分离效果。用胰蛋白酶将上皮组织消化成细胞悬液,台盼蓝染色法检测细胞的活性。结果 翻转-固定分离法分离的结膜上皮菲薄,光镜下观察结膜上皮下无纤维组织残留。台盼蓝染色法证实上皮细胞活性良好。结论 翻转-固定分离法是一种可靠、廉价的获取单纯结膜上皮的方法。获取的结膜上皮可用于进一步的细胞培养。

A new technique was been investigated to obtain intact, pure sheets of rabbit conjunctival epithelium for the culture of con-junctival epithelial cells. Methods Conjunctival epithelium and matrix were isolated by the method of overturn - fixing. Freed conjunctival epithelium sheet was fixed, dehydrated, embeded, cut and stained with hematoxylin and eosin and examined by light microscopy. Trypain was to digest epithelium. The ratio of live cell was determined by trypan blue staining. Results Freed conjunctival epithelium sheet was extremely thin. There were no stromal elements left under conjunctival epithelium. Ratio of live cell was 98% . Conclusion Method of overturn - fixing was a cheap and reliable technique for isolating conjunctival epithelium. With the technique, pure population of intact conjunctival epithelium can be obtained for further tissue culture.