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Gene Expression Profiles and Retinal Potential of Stem/Progenitor Cells Derived from Human Iris and Ciliary Pigment Epithelium

机译:人虹膜和睫状色素上皮细胞衍生的干/祖细胞的基因表达谱和视网膜电位

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The aim of our study was to isolate and characterize the properties of neurospheres and differentiated cellular progeny derived from iris and ciliary pigment epithelial (IPE and CPE) cells of human cadaveric eyes. In this study we investigated the gene expression profiles of the stem/progenitor cells and the differentiated progeny derived from IPE and CPE cells, as the changes underlying differentiation of the stem/progenitor derived from the IPE and CPE cells from human cadaveric eye are essentially unknown. The IPE and CPE cells from human cadaver eyes were cultured in the presence of mitogens to generate neurospheres (NS) and the growth characteristics were evaluated. The Neurospheres (NS) were plated under conditions inducing differentiation and their potential was analyzed by RT-PCR, immunocytochemistry, calcium imaging studies and microarray studies to measure the changes involved in the process of differentiation. The IPE and CPE cells can generate NS containing progenitor cells in the presence of mitogens and were capable of producing different retinal cell types as demonstrated by RT-PCR and immunocytochemistry. The cluster analyses of the differentially expressed genes show the dynamic changes that occur during the course of IPE and CPE neurospheres differentiating into retinal progeny. The study gives clues towards the changes that occur during differentiation from NS into retinal progeny. In the present study we have demonstrated the expansion and maintenance of SCs from IPE and CPE of cadaveric eyes. These cells maintain their self-renewal properties and the ability to differentiate along retinal cell lineages and hence could be a practical source of donor cells for ex-vivo stem cell therapy during retinal degeneration.
机译:我们研究的目的是分离和表征人尸体眼睛的虹膜和睫状色素上皮(IPE和CPE)细胞衍生的神经球和分化的细胞后代的特性。在这项研究中,我们研究了干/祖细胞以及来自IPE和CPE细胞的分化后代的基因表达谱,因为人类尸体眼睛的IPE和CPE细胞衍生的干/祖细胞分化的根本变化是未知的。在有丝分裂原的存在下培养人尸眼的IPE和CPE细胞,生成神经球(NS),并评估其生长特性。将神经球(NS)置于诱导分化的条件下进行铺板,并通过RT-PCR,免疫细胞化学,钙成像研究和微阵列研究分析其潜力,以测量分化过程中涉及的变化。 IPE和CPE细胞可以在有丝分裂原存在的情况下产生含NS的祖细胞,并能够产生不同的视网膜细胞类型,如RT-PCR和免疫细胞化学所证实。差异表达基因的聚类分析显示在IPE和CPE神经球分化为视网膜后代的过程中发生的动态变化。该研究为从NS分化为视网膜后代的过程中发生的变化提供了线索。在本研究中,我们已经证明了尸体眼睛的IPE和CPE对SC的扩展和维持。这些细胞保持其自我更新特性和沿视网膜细胞谱系分化的能力,因此可能是视网膜变性期间离体干细胞治疗的实际供体细胞来源。

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