Facile isolation and the characterization of human retinal stem cells
Facile isolation and the characterization of human retinal stem cells
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DOI：
10.1073/pnas.0401596101
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发表时间：
2004-11-02
影响因子：
11.1
通讯作者：
van der Kooy, D
中科院分区：
文献类型：
--
作者：
Coles, BLK;Angénieux, B;van der Kooy, D
这项研究鉴定和表征了出生后早期至70岁人眼的视网膜干细胞(RSCs)。分离人眼不同亚区，用克隆球体形成法进行培养。干细胞仅来自视网膜睫状缘的折叠部和平坦部，频率约为1：500。为了检测长期的自我更新，采用球法和单层传代两种方法。通过单球传代实验，原始球被解离并复制，单个球表现出100%的自我更新，在随后的每一代中，单个球产生一个或多个新的球。在分化条件下进行视网膜克隆球的培养，以检测其后代的分化潜能。这些球体产生了所有不同类型的视网膜细胞，显示出多潜能。因此，人眼含有少量具有视网膜干细胞特征(增殖、自我更新和多潜能)的细胞(每只眼大约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状细胞移植到出生后第1天的NCD/SCID小鼠和胚胎鸡眼中。RSCs的后代能够存活、迁移、整合和分化到神经视网膜，特别是作为光感受器。它们易于分离、整合和分化，提示人类视网膜干细胞最终可能在治疗人类视网膜疾病方面有价值。
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 approximate to1:500. 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 (approximate to10,000 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 NCD/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.