Human Retinal Progenitor Cells

描述(申请人提供)：我们已分离出具有人类视网膜祖细胞特征的克隆细胞系。这些细胞来自视网膜母细胞瘤肿瘤。虽然肿瘤细胞在R81基因上是突变的，但祖细胞没有表现出突变(除非天生携带在基因组中)。这里提出的研究涉及完全野生型的单克隆细胞系。 要完全符合视网膜祖细胞的资格，细胞必须是自我更新的，必须具有祖细胞的标志，并且必须具有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，表达神经和视网膜前体细胞特有的标志，并在移植后呈现视网膜细胞的形态特征。我们的目标是(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。研究建议将细胞系置于促进其分化的环境中，包括改变培养液中的生长因子，与胚胎或出生后的大鼠视网膜细胞重组培养，将细胞引入出生后早期的大鼠或小鼠视网膜外植体，以及将细胞移植到发育中的大鼠视网膜。祖细胞将被标记增强的绿色荧光蛋白，并将评估它们表达视网膜标志物和形成合适的视网膜细胞形态的潜力。此外，我们建议(2)为将来的研究提供更多的细胞系以建立细胞库。 充分鉴定的祖细胞为研究视网膜分化提供了宝贵的资源，可能为退化或受损的哺乳动物视网膜提供细胞替代资源，并为预防或修复病变视网膜的药物提供资源。

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.