Human Retinal Progenitor Cells

描述（由申请人提供）：我们分离了显示人视网膜祖细胞特征的克隆细胞系。这些细胞来源于视网膜母细胞瘤肿瘤。虽然肿瘤细胞在R81基因中是突变的，但祖细胞不显示突变（除非基因组中固有携带）。这里提出的研究涉及完全野生型单克隆细胞系。 为了完全符合视网膜祖细胞的资格，细胞必须自我更新，必须显示祖细胞的标记，并且必须显示分化为视网膜细胞的潜力。初步研究表明，这些细胞是自我更新的，表达神经，视网膜祖细胞的特征性标志物，并在移植后呈现视网膜细胞的形态特征。我们的目的是（1）获得这些细胞具有分化为功能性视网膜细胞的能力的证据。研究提出了将细胞系置于将引起其分化的环境中，包括改变培养基中的生长因子，用胚胎或出生后大鼠视网膜细胞进行再聚集培养，将细胞引入出生后早期大鼠或小鼠视网膜外植体中，以及将细胞移植到发育中的出生后大鼠视网膜中。将用增强的绿色荧光蛋白标记祖细胞，并评估其表达视网膜标记物和形成视网膜细胞的适当形态的潜力。此外，我们建议（2）获得额外的细胞系，以储存细胞用于未来的研究。 完全表征的祖细胞为视网膜分化的研究提供了宝贵的资源，可以为退化或受损的哺乳动物视网膜中的细胞替代提供资源，并为递送试剂以预防或修复患病视网膜提供资源。

DESCRIPTION (provided by applicant): We have isolated clonal cell lines that show traits of human retinal progenitor cells. The cells are derived from retinoblastoma tumors. While the tumor cells are mutant in the R81 gene, the progenitors do not show mutations (unless inherently carried in the genome). Studies proposed here deal with fully wildtype monoclonal cell lines. To fully qualify as retinal progenitors, the cells must be self renewing, must show markers of progenitor cells, and must show the potential to differentiate into retinal cells. Preliminary studies show that these cells are self renewing, express markers characteristic of neural, retinal progenitor cells and assume morphological traits of retinal cells upon transplantation. Our objective here is (1) to obtain evidence that these cells have the capacity to differentiate into functional retinal cells. Studies are proposed to subject the cell lines to environments that will provoke their differentiation, including altering growth factors in culture media, reaggregation culture with embryonic or postnatal rat retinal cells, introduction of cells into early postnatal rat or mouse retinal explants, and transplantation of cells into developing postnatal rat retina. Progenitor cells will be marked with enhanced green fluorescent protein and their potential to express retinal markers and develop appropriate morphology of retinal cells will be assessed. In addition, we propose (2) to derive additional cell lines to bank cells for future studies. Fully characterized progenitor cells provide an invaluable resource for the study of retinal differentiation, may provide a resource for cell replacement in degenerated or damaged mammalian retina, and provide a resource for delivery of agents to prevent or repair diseased retina.