Defoe D M, Easterling K C
Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta 30912-2000.
Invest Ophthalmol Vis Sci. 1994 Apr;35(5):2466-76.
The authors aimed to establish an in vitro model of the retinal pigment epithelium (RPE) suitable for studies of neural retina-epithelium interactions.
Sheets of RPE were obtained from Xenopus laevis eyes by Dispase treatment of intact globes. After dispersal in trypsin-EDTA solution, cells were plated onto Matrigel-coated microporous membrane filters and grown in a serum-free defined medium.
Confluent cell monolayers obtained after 7 to 10 days of culture appeared by electron microscopy to be morphologically polarized, established junctional complexes, and exhibited transepithelial resistances ranging from 300 to 500 omega.cm2. As did the native epithelium, these cells formed cytoskeletons consisting of circumferential bands of actin myofilaments at their periphery and whorls of cytokeratin intermediate filaments throughout the cytoplasm. Co-culture of freshly isolated neural retinas with monolayers resulted in the apparent reattachment of photoreceptors to the RPE within 3 hours.
Primary cultures of amphibian RPE that express phenotypic characteristics of the epithelium in situ are also capable of establishing adhesive interactions with isolated neural retinas.
作者旨在建立一种适用于研究神经视网膜 - 上皮细胞相互作用的视网膜色素上皮（RPE）体外模型。
通过对完整眼球进行Dispase处理，从非洲爪蟾眼中获取RPE片层。在胰蛋白酶 - EDTA溶液中分散后，将细胞接种到基质胶包被的微孔膜滤器上，并在无血清限定培养基中培养。
培养7至10天后获得的汇合细胞单层经电子显微镜观察显示形态极化，形成连接复合体，跨上皮电阻范围为300至500Ω·cm²。与天然上皮一样，这些细胞形成细胞骨架，其周边由肌动蛋白肌丝的圆周带组成，整个细胞质中有细胞角蛋白中间丝的螺旋。新鲜分离的神经视网膜与单层细胞共培养导致光感受器在3小时内明显重新附着于RPE。
原位表达上皮细胞表型特征的两栖类RPE原代培养物也能够与分离的神经视网膜建立黏附相互作用。
