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目的 探索羊膜基底膜对结膜上皮细胞形态功能的影响。方法 把“去上皮”羊膜按照基底膜或实质面向上分为两组,接种结膜上皮细胞,同样条件下培养。取培养14天的两组标本进行组织学(HE和PAS染色),透射电镜,以及免疫组化检查(包括抗角蛋白和抗波形蛋白染色)。结果细胞72小时内贴附在羊膜上。基底膜面接种的上皮细胞顶端见微绒毛,单层排列,细胞间有丰富桥粒,基底膜完整,可见半桥粒,抗角蛋白染色阳性,抗波形蛋白染色阴性;实质面接种的上皮细胞浆内有大量脂滴,表面未见微绒毛,细胞松散堆积,与羊膜连接不紧密,未见桥粒和半桥粒,抗角蛋白染色弱阳性,抗波形蛋白染色强阳性。结论 羊膜基底膜是结膜上皮细胞体外培养的良好载体,有利于上皮细胞生长、黏附、分化,并能减缓上皮细胞老化。
Objective To explore the effect of amniotic basement membrane on the morphological function of conjunctival epithelial cells. Methods The “de-epithelial” amniotic membrane was divided into two groups according to the basement membrane or the parenchyma. The conjunctival epithelial cells were inoculated and cultivated under the same conditions. Histology (HE and PAS staining), transmission electron microscopy, and immunohistochemistry (including anti-keratin and anti-vimentin staining) were performed on two groups of 14-day culture specimens. As a result, the cells adhered to the amniotic membrane within 72 hours. The top of epithelial cells inoculated on the surface of basal membrane were seeded with microvilli and arranged in a single layer with plentiful desmosomes. The basal membrane was intact. The hemidesmosomes, positive anti-keratin staining and negative anti-vimentin staining showed that the epithelial cells There was a large number of lipid droplets in the pulp, no microvilli on the surface, loosely packed cells, and the amniotic membrane were not closely connected. No desmosomes and hemidesmosomes were found. Anti-keratin staining was weakly positive and anti-vimentin staining was strongly positive. Conclusion Amniotic basement membrane is a good carrier for cultured conjunctival epithelial cells in vitro, which is conducive to epithelial cell growth, adhesion, differentiation, and can slow the aging of epithelial cells.