"去上-皮"羊膜對體外培養的結膜上皮細胞的作用

目的探索羊膜基底膜對結膜上皮細胞形態功能的影響.方法把"去上皮"羊膜按照基底膜或實質面向上分為兩組,接種結膜上皮細胞,同樣條件下培養.取培養14天的兩組標本進行組織學(HE和PAS染色),透射電鏡,以及免疫組化檢查(包括抗角蛋白和抗波形蛋白染色).結果細胞72小時内貼附在羊膜上.基底膜面接種的上皮細胞頂端見微絨毛,單層排列,細胞間有豐富橋粒,基底膜完整,可見半橋粒,抗角蛋白染色陽性,抗波形蛋白染色陰性;實質面接種的上皮細胞漿内有大量脂滴,表面未見微絨毛,細胞鬆散堆積,與羊膜連接不緊密,未見橋粒和半橋粒,抗角蛋白染色弱陽性,抗波形蛋白染色强陽性.結論羊膜基底膜是結膜上皮細胞體外培養的良好載體,有利于上皮細胞生長、黏附、分化,并能减緩上皮細胞老化.

The effects of acellular human amniotic membrane on cultured human conjunctival epithelium cells

Objective To study the influence of amniotic membrane on the morphology and function of human conjuncitival epithelium cells. Methods Isolated human conjunctival epithelium cells were cultured on the basement membrane and stromal surface of an acellular human amniotic membrane. After 14 days, the cultured epithelium cells were examined histologically with staining of HE, PAS, transmission electronic microscopy, and immunohistochemically using monoclonal antibodies, Pan-Keration and Vimentin. Results The cultured cells attached to the amniotic membrane within 72 hours. Conjunctival epithelium cells cultured on basement membrane formed a well-organized monolayer. When cultured on the stromal side, they formed clumps and appeared less ordered. Transmission election microscopy showed cultures on basement membrane had more mitochondria and endoplasmic reticulum than cultures on stromal. There were a large number of intracellular desmosomes and hemidesmosomes in the cells cultured on basement membrane, while no one was observed in the cells cultured on stromal. On immunohistochemical examination, cultures on basement membrane were stained positive for keratin, but cultures on stromal were stained positive for vimentin. Conclusion The basement membrane of amniotic membrane is the proper substrate for the growth of human conjunctival epithelium cells. It facilitates epithelial proliferation, prevents epithelial aging, and reinforces adhesion of the epithelial cells. It can guide the epithelial cells differentiation along the right direction.