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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Facile isolation and the characterization of human retinal stem cells.
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Facile isolation and the characterization of human retinal stem cells.

机译：易于分离和人类视网膜干细胞的表征。

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This study identifies and characterizes retinal stem cells (RSCs) in early postnatal to seventh-decade human eyes. Different subregions of human eyes were dissociated and cultured by using a clonal sphere-forming assay. The stem cells were derived only from the pars plicata and pars plana of the retinal ciliary margin, at a frequency of approximately 1:500. To test for long-term self-renewal, both the sphere assay and monolayer passaging were used. By using the single sphere passaging assay, primary spheres were dissociated and replated, and individual spheres demonstrated 100% self-renewal, with single spheres giving rise to one or more new spheres in each subsequent passage. The clonal retinal spheres were plated under differentiation conditions to assay the differentiation potential of their progeny. The spheres were produced all of the different retinal cell types, demonstrating multipotentiality. Therefore, the human eye contains a small population of cells ( approximately 10,000 cells per eye) that have retinal stem-cell characteristics (proliferation, self-renewal, and multipotentiality). To test the in vivo potential of the stem cells and their progeny, we transplanted dissociated human retinal sphere cells, containing both stem cells and progenitors, into the eyes of postnatal day 1 NOD/SCID mice and embryonic chick eyes. The progeny of the RSCs were able to survive, migrate, integrate, and differentiate into the neural retina, especially as photoreceptors. Their facile isolation, integration, and differentiation suggest that human RSCs eventually may be valuable in treating human retinal diseases.
机译：这项研究鉴定并鉴定了出生后早期至七十年代人眼中的视网膜干细胞（RSC）。使用克隆球形成测定法分离并培养人眼的不同亚区域。干细胞仅以大约1：500的频率来自视网膜睫状缘的腮腺和睫状突。为了测试长期自我更新，使用了球形测定法和单层传代法。通过使用单球通过测试，主球被解离和重新填充，单个球表现出100％的自我更新，单个球在随后的每个传球中都会产生一个或多个新球。在分化条件下接种克隆性视网膜球，以测定其子代的分化潜能。这些球是由所有不同的视网膜细胞类型产生的，显示出多能性。因此，人眼包含少量具有视网膜干细胞特征（增殖，自我更新和多能性）的细胞（每只眼睛约10,000个细胞）。为了测试干细胞及其后代的体内潜能，我们将含有干细胞和祖细胞的分离的人视网膜球细胞移植到了出生后第1天的NOD / SCID小鼠的眼睛和胚胎鸡眼中。 RSC的后代能够存活，迁移，整合并分化为神经视网膜，尤其是作为感光细胞。它们的简便隔离，整合和分化表明，人类RSC最终在治疗人类视网膜疾病中可能有价值。

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