Tsai R J, Ho Y S, Chen J K
Department of Ophthalmology, Chang Gung Memorial Hospital, Linko, Taoyuan, Taiwan.
Invest Ophthalmol Vis Sci. 1994 May;35(6):2865-75.
To study the effects of collagen matrix and fibroblasts on the growth and development of human bulbar conjunctival epithelial cells.
Human bulbar conjunctival epithelial cells were cultured on three-dimensional collagen gels containing either normal human conjunctival fibroblasts (HCF), Swiss 3T3 cells, or no cells. After 1 week of culturing, half of the cultures were raised to the air-liquid interface and the rest of the cultures remained submerged. On day 14, cultures were fixed and sectioned for light and electron microscopic studies.
Conjunctival epithelial cells cultured on fibroblast-contracted collagen lattice developed into a multicell-layer epithelium with characteristic epithelial structural features including microvilli, desmosomes, early hemidesmosomes, and basement membrane-like structures. Formation of all or some of the above features appeared to be influenced by the type of fibroblasts in the collagen lattices. Structures such as hemidesmosomes and basement membrane were only observed in epithelium developed on 3T3- but not on conjunctival fibroblast-condensed collagen lattices. In contrast, goblet cell differentiation was only observed in epithelia developed on normal HCF-supported collagen matrix. Epithelial cells cultured on acellular collagen gels did not develop into multicell-layer epithelium, and no differentiated characteristics were observed.
These results indicate that the type of fibroblasts dispersed in the collagen matrix plays an important role in the development and differentiation of conjunctival epithelial cells. Normal HCF-dispersed collagen matrix was less growth stimulating to epithelial cells and allowed them to undergo goblet cell differentiation. In contrast, 3T3-dispersed collagen matrix was more growth stimulating, resulting in thicker epithelium with a higher degree of stratification.
研究胶原基质和成纤维细胞对人球结膜上皮细胞生长发育的影响。
将人球结膜上皮细胞培养在含有正常人结膜成纤维细胞（HCF）、瑞士3T3细胞或无细胞的三维胶原凝胶上。培养1周后，将一半培养物提升至气液界面，其余培养物保持浸没状态。在第14天，将培养物固定并切片进行光镜和电镜研究。
在成纤维细胞收缩的胶原晶格上培养的结膜上皮细胞发育成多细胞层上皮，具有特征性的上皮结构特征，包括微绒毛、桥粒、早期半桥粒和基底膜样结构。上述全部或部分特征的形成似乎受胶原晶格中成纤维细胞类型的影响。半桥粒和基底膜等结构仅在3T3细胞收缩的胶原晶格上发育的上皮中观察到，而在结膜成纤维细胞收缩的胶原晶格上未观察到。相反，杯状细胞分化仅在正常HCF支持的胶原基质上发育的上皮中观察到。在无细胞胶原凝胶上培养的上皮细胞未发育成多细胞层上皮，也未观察到分化特征。
这些结果表明，分散在胶原基质中的成纤维细胞类型在结膜上皮细胞的发育和分化中起重要作用。正常HCF分散的胶原基质对上皮细胞的生长刺激较小，使其能够进行杯状细胞分化。相比之下，3T3分散的胶原基质对生长的刺激更大，导致上皮更厚，分层程度更高。
