The effects of amniotic membrane on retinal pigment epithelial cell differentiation.
The effects of amniotic membrane on retinal pigment epithelial cell differentiation.
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DOI：
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发表时间：
2005-01
期刊：
影响因子：
2.2
通讯作者：
K. Ohno-Matsui;S. Ichinose;K. Nakahama;Takeshi Yoshida;A. Kojima;M. Mochizuki;I. Morita
中科院分区：
文献类型：
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作者：
K. Ohno-Matsui;S. Ichinose;K. Nakahama;Takeshi Yoshida;A. Kojima;M. Mochizuki;I. Morita
目的 检测羊膜（AM）上培养的视网膜色素上皮（RPE）细胞的特性。本研究探讨了 AM 如何调节 RPE 细胞分化。方法 人 RPE 细胞在分散酶处理过的 AM 的基底膜侧培养。细胞汇合一周后，终止培养，收集条件培养基，并提取总RNA。对被认为参与分化 RPE 功能的几个基因的表达模式进行了评估。通过透射电子显微镜评估超微结构的变化。结果在形态学上，AM 培养的 RPE 细胞表现出超微结构上皮特征，例如顶膜微绒毛和细胞间连接。与塑料上培养的细胞相比，AM 上培养的 RPE 细胞中 RPE65、CRALBP、bestropin 和酪氨酸酶相关蛋白 (TRP)-2 的基因表达上调。此外，在AM上培养的细胞中，血管内皮生长因子、血小板反应蛋白-1和色素上皮衍生因子的蛋白质产量显着增加。然而，组织蛋白酶 D、脑源性神经营养因子和碱性成纤维细胞生长因子的基因表达在塑料或 AM 上培养的 RPE 细胞之间没有差异。结论 AM 培养的 RPE 细胞在形态学上表现出上皮表型，并且对维持视网膜稳态重要的几种生长因子上调。 AM 可能是一种有用的基质基质，可保留 RPE 的分化和上皮表型，用于视网膜下移植。
PURPOSE To examine the characteristics of retinal pigment epithelial (RPE) cells cultured on amniotic membrane (AM). The present study examined how AM modulates RPE cell differentiation. METHODS Human RPE cells were cultured on the basement membrane side of dispase treated AM. After one week of cellular confluence, cultures were terminated, conditioned medium was collected, and total RNA was extracted. The expression pattern of several genes considered to participate in the function of differentiated RPE was evaluated. Ultrastructural changes were evaluated by transmission electron microscopy. RESULTS Morphologically, RPE cells cultured on AM exhibited ultrastructural epithelial features such as microvilli of the apical membrane and intercellular junctions. Gene expression of RPE65, CRALBP, bestrophin, and tyrosinase related protein (TRP)-2 was upregulated in RPE cells cultured on AM compared to cells cultured on plastic. In addition, protein production of vascular endothelial growth factor, thrombospondin-1, and pigment epithelium derived factor was markedly increased in cells cultivated on AM. Gene expression of cathepsin D, brain derived neurotrophic factor, and basic fibroblast growth factor, however, did not differ between RPE cells cultured on plastic or AM. CONCLUSIONS RPE cells cultured on AM demonstrated an epithelial phenotype morphologically and several growth factors important for maintaining retinal homeostasis were upregulated. AM might be a useful matrix substrate to retain the differentiated and epithelial phenotype of RPE for subretinal transplantation.