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首页> 外文期刊>Investigative ophthalmology & visual science >In Vitro Comparison Of Structural And Functional Characteristics Of Cultured Autologous Monolayer Sheets Of Iris Pigmentary Epithelial And Retinal Pigmentary Epithelial Cells
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In Vitro Comparison Of Structural And Functional Characteristics Of Cultured Autologous Monolayer Sheets Of Iris Pigmentary Epithelial And Retinal Pigmentary Epithelial Cells

机译：虹膜色素上皮细胞和视网膜色素上皮细胞自体培养单层片的结构和功能特征的体外比较

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Purpose: : The purpose of this study is to culture and establish viable autologous monolayer sheets of sheep IPE cells and RPE cells in vitro. Second, we will compare the morphology and functional aspects of the IPE and RPE cells on the monolayer sheets to ascertain the preservation of their functions. Methods: : Posterior IPE and RPE cells were gently scraped out of freshly enucleated sheep eyes following established protocols. Equal number of cells were seeded on pretreated human amniotic membrane and incubated for 10 days in DMEM medium supplemented with 10% FBS and Pen/Strep. Histological and immunological studies were performed on paraffin embedded sections to study the different morphological features of the cells. Phagocytosis was assayed by fluorescent microscopy after daily feeding the cells with FITC-conjugated oxidized bovine rod outer segments (1x106/ml) for 10 hours and then for 24, 48 and 72 hours. Results: : Amniotic membrane acts as a robust matrix for monolayer cell cultivation with cells retaining native characteristics. Both IPE and RPE cells stained positive with cytokeratin confirming their epithelial origin. IPE cells propagated similarly like RPE cells on the amniotic membrane forming monolayer sheets and in fact maintained cell to cell junctions. In addition, on the amniotic membrane, IPE cells depicted melanin granules and structural similarity to the isolated RPE cells and as seen on hematoxylin-eosin sections. Functionally, IPE cells were able to phagocyte FITC labeled rod outer segments with 20% lower efficiency in comparison to RPE cells at 10 and 24 hour incubation period. However, at 48 and 72 hour time points IPE cells showed higher accumulates of ROS than RPE cells. Conclusions: : TIPE cells can be grown as monolayer sheets on amniotic membrane and they resemble not only structural characteristics but also physiological properties with RPE cells in vitro. Therefore, IPE cells are ideal cells for RPE cell replacement especially when grown as monolayer sheets on amniotic membrane. This holds strong clinical significance with respect to therapeutic approaches in treatment of severe dry and exudative macular degeneration.
机译：目的：：本研究的目的是体外培养和建立绵羊IPE细胞和RPE细胞的自体单层活板。其次，我们将比较单层纸上IPE和RPE细胞的形态和功能方面，以确定其功能是否得以保留。方法：按照既定规程从新去核的绵羊眼中轻轻刮除后部IPE和RPE细胞。将等量的细胞接种在预处理的人羊膜上，并在补充有10％FBS和Pen / Strep的DMEM培养基中孵育10天。对石蜡包埋的切片进行了组织学和免疫学研究，以研究细胞的不同形态特征。每天用FITC偶联的氧化牛杆外段（1x106 / ml）喂食细胞10小时，然后持续24、48和72小时，然后通过荧光显微镜检查吞噬作用。结果：羊膜可作为单层细胞培养的坚固基质，同时保留天然特性。 IPE和RPE细胞均被细胞角蛋白染色阳性，证实了它们的上皮起源。 IPE细胞像RPE细胞一样在羊膜上形成单层薄片，但实际上维持了细胞与细胞的连接。此外，在羊膜上，IPE细胞描绘了黑色素颗粒，并且与分离的RPE细胞具有结构相似性，并且在苏木精-曙红切片中也可以看到。在功能上，与RPE细胞在10和24小时的孵育期相比，IPE细胞能够吞噬细胞FITC标记的杆外部片段，效率降低了20％。但是，在48小时和72小时的时间点，IPE细胞显示的ROS积累量高于RPE细胞。结论：TIPE细胞可以在羊膜上单层生长，并且不仅具有RPE细胞的结构特征，而且还具有生理特性。因此，IPE细胞是替代RPE细胞的理想细胞，尤其是当在羊膜上单层生长时。就严重干燥和渗出性黄斑变性的治疗方法而言，这具有重要的临床意义。

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