当前位置: X-MOL 学术Stem Cell Res. Ther. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Protective effects of human iPS-derived retinal pigmented epithelial cells on retinal degenerative disease.
Stem Cell Research & Therapy ( IF 7.1 ) Pub Date : 2020-03-04 , DOI: 10.1186/s13287-020-01608-8
Deliang Zhu 1, 2 , Mengyuan Xie 1 , Fabian Gademann 2 , Jixing Cao 3 , Peiyuan Wang 3 , Yonglong Guo 2 , Lan Zhang 3 , Ting Su 3 , Jun Zhang 1 , Jiansu Chen 2, 3, 4
Affiliation  

Retinitis pigmentosa (RP) is an inherited retinal disease characterized by progressive loss of photoreceptor cells. This study aim at exploring the effect of retinal pigment epithelium (RPE) derived from human-induced pluripotent stem cell (hiPSC-RPE) on the retina of retinal degeneration 10 (rd10) mice, which are characterized with progressive photoreceptor death. We generated RPE from hiPSCs by sequential supplementation with retinal-inducing factors and RPE specification signaling factors. The three-dimensional (3D) spheroid culture method was used to obtain optimal injectable hiPSC-RPE cells. Subretinal space transplantation was conducted to deliver hiPSC-RPE cells into the retina of rd10 mice. Neurotrophic factor secretion from transplanted hiPSC-RPE cells was detected by enzyme-linked immunosorbent assay (ELISA). Immunostaining, Western blotting, electroretinography (ERG), and visual behavior testing were performed to determine the effects of hiPSC-RPE on the retinal visual function in rd10 mice. Our data demonstrated that hiPSC-RPE cells exhibited classic RPE properties and phenotype after the sequential RPE induction from hiPSCs. hiPSC-RPE cells co-cultured with mouse retinal explants or retinal ganglion cells 5 (RGC5) exhibited decreased apoptosis. The viability and functional properties of hiPSC-RPE cells were enhanced by 3D spheroid culture. Transplanted hiPSC-derived RPE cells were identified by immunostaining with human nuclear antigen staining in the retina of rd10 14 days after subretinal space injection. The pigment epithelium-derived factor level was increased significantly. The expression of CD68, microglial activation marker, reduced after transplantation. The light avoidance behavior and ERG visual function in rd10 mice improved by the transplantation of hiPSC-RPE cells. Our findings suggest that injectable hiPSC-RPE cells after 3D spheroid culture can rescue the structure and function of photoreceptors by sub-retinal transplantation, which lay the foundation for future clinical cell therapy to treat RP and other retinal degeneration diseases.

中文翻译:

人 iPS 衍生的视网膜色素上皮细胞对视网膜退行性疾病的保护作用。

色素性视网膜炎 (RP) 是一种遗传性视网膜疾病,其特征是感光细胞进行性丧失。本研究旨在探索人诱导多能干细胞 (hiPSC-RPE) 衍生的视网膜色素上皮 (RPE) 对视网膜变性 10 (rd10) 小鼠视网膜的影响,这些小鼠以进行性光感受器死亡为特征。我们通过连续补充视网膜诱导因子和 RPE 规范信号因子从 hiPSC 生成 RPE。三维 (3D) 球体培养方法用于获得最佳可注射 hiPSC-RPE 细胞。进行视网膜下间隙移植以将hiPSC-RPE细胞递送到rd10小鼠的视网膜中。通过酶联免疫吸附试验(ELISA)检测移植的hiPSC-RPE细胞的神经营养因子分泌。免疫染色,进行蛋白质印迹、视网膜电图 (ERG) 和视觉行为测试以确定 hiPSC-RPE 对 rd10 小鼠视网膜视觉功能的影响。我们的数据表明,hiPSC-RPE 细胞在从 hiPSC 连续诱导 RPE 后表现出经典的 RPE 特性和表型。与小鼠视网膜外植体或视网膜神经节细胞 5 (RGC5) 共培养的 hiPSC-RPE 细胞表现出细胞凋亡减少。3D 球体培养增强了 hiPSC-RPE 细胞的活力和功能特性。移植的hiPSC衍生的RPE细胞通过在视网膜下腔注射14天后rd10的视网膜中用人核抗原染色进行免疫染色来鉴定。色素上皮衍生因子水平显着增加。移植后小胶质细胞活化标志物CD68的表达降低。移植hiPSC-RPE细胞后rd10小鼠的避光行为和ERG视觉功能得到改善。我们的研究结果表明,3D 球体培养后可注射的 hiPSC-RPE 细胞可以通过视网膜下移植来拯救光感受器的结构和功能,这为未来临床细胞疗法治疗 RP 和其他视网膜变性疾病奠定了基础。
更新日期:2020-03-04
down
wechat
bug