Awata T, Nishida T, Nakagawa S, Manabe R
Department of Ophthalmology, Osaka University Medical School, Japan.
Jpn J Ophthalmol. 1989;33(1):132-43.
The production of fibronectin (FN) and its response to serum or epidermal growth factor (EGF) were investigated in three different types of rabbit corneal cells cultured in vitro. The corneal epithelial cells, stromal fibroblasts (keratocytes) and endothelial cells were separately cultured in different media: basic medium containing minimal serum (0.5%), basic medium with supplementary serum at a final concentration of 10% and basic medium with 100 ng/ml EGF, respectively. FN production by each type of cell was examined either by the immunofluorescent staining method or by the metabolic labeling method followed by immunoprecipitation of FN in the culture medium. Each type of corneal cell produced and secreted FN. FN secretion into the culture medium by keratocytes and by endothelial cells was enhanced by the addition of EGF. However, FN secretion by epithelial cells was lowered by the additional serum or EGF. Furthermore, when the epithelial cells were cultured in the basic medium, DNA synthesis was low but FN secretion was high. These results suggest that the control mechanism of FN production differs between epithelial cells and keratocytes or endothelial cells.
研究了体外培养的三种不同类型兔角膜细胞中纤连蛋白(FN)的产生及其对血清或表皮生长因子(EGF)的反应。角膜上皮细胞、基质成纤维细胞(角膜细胞)和内皮细胞分别在不同培养基中培养:分别为含最低血清(0.5%)的基础培养基、最终浓度为10%补充血清的基础培养基以及含100 ng/ml EGF的基础培养基。通过免疫荧光染色法或代谢标记法,随后对培养基中的FN进行免疫沉淀,检测每种细胞类型的FN产生情况。每种角膜细胞都产生并分泌FN。添加EGF可增强角膜细胞和内皮细胞向培养基中分泌FN。然而,额外的血清或EGF会降低上皮细胞的FN分泌。此外,当上皮细胞在基础培养基中培养时,DNA合成较低,但FN分泌较高。这些结果表明,上皮细胞与角膜细胞或内皮细胞之间FN产生的调控机制不同。