Department of Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, London, UK.
Biomaterials. 2010 Oct;31(30):7726-37. doi: 10.1016/j.biomaterials.2010.07.012. Epub 2010 Jul 31.
We describe, for the first time, the use of cellular plastic compressed collagen as a substrate for human limbal epithelial cell expansion and stratification. The characteristics of expanded limbal epithelial cells on either acellular collagen constructs or those containing human limbal fibroblasts were compared to a human central cornea control. After compression, human fibroblasts in collagen constructs remained viable and limbal epithelial cells were successfully expanded on the surface. After airlifting, a multilayered epithelium formed with epithelial cell morphology very similar to that of cells in the central cornea. Immunochemical staining revealed expression of basement membrane proteins and differentiated epithelial cell markers found in native central cornea. Ultrastructural analysis revealed cells on collagen constructs had many features similar to central cornea, including polygonal, tightly opposed surface epithelial cells with microvilli and numerous desmosomes at cell-cell junctions. Taken together, these data demonstrate that plastic compressed collagen constructs can form the basis of a biomimetic tissue model for in vitro testing and could potentially provide a suitable alternative to amniotic membrane as a substrate for limbal epithelial cell transplantation.
我们首次描述了细胞塑料压缩胶原作为人角膜缘上皮细胞扩增和分层的基质的用途。将扩增的角膜缘上皮细胞在脱细胞胶原构建体上或包含人角膜缘成纤维细胞的构建体上的特征与人类中央角膜对照进行了比较。压缩后，胶原构建体中的人成纤维细胞保持存活，并且角膜缘上皮细胞成功地在表面上扩展。气浮后，形成了具有与中央角膜中细胞非常相似的上皮细胞形态的多层上皮。免疫组织化学染色显示出在天然中央角膜中发现的基底膜蛋白和分化的上皮细胞标志物的表达。超微结构分析显示，胶原构建体上的细胞具有许多与中央角膜相似的特征，包括多边形、紧密相对的表面上皮细胞，具有微绒毛和细胞-细胞连接处的许多桥粒。综上所述，这些数据表明塑料压缩胶原构建体可以作为体外测试的仿生组织模型的基础，并可能为角膜缘上皮细胞移植提供合适的替代羊膜的基质。
