Thomas Biju B, Zhu Danhong, Zhang Li, Thomas Padmaja B, Hu Yuntao, Nazari Hossein, Stefanini Francisco, Falabella Paulo, Clegg Dennis O, Hinton David R, Humayun Mark S
Department of Ophthalmology, USC Roski Eye Institute, University of Southern California, Los Angeles, California, United States 2USC Institute for Biomedical Therapeutics, University of Southern California, Los Angeles, California, United States.
Department of Ophthalmology, USC Roski Eye Institute, University of Southern California, Los Angeles, California, United States 3Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, United States.
Invest Ophthalmol Vis Sci. 2016 May 1;57(6):2877-87. doi: 10.1167/iovs.16-19238.
To determine the safety, survival, and functionality of human embryonic stem cell-derived RPE (hESC-RPE) cells seeded on a polymeric substrate (rCPCB-RPE1 implant) and implanted into the subretinal (SR) space of Royal College of Surgeons (RCS) rats.
Monolayers of hESC-RPE cells cultured on parylene membrane were transplanted into the SR space of 4-week-old RCS rats. Group 1 (n = 46) received vitronectin-coated parylene membrane without cells (rMSPM+VN), group 2 (n = 59) received rCPCB-RPE1 implants, and group 3 (n = 13) served as the control group. Animals that are selected based on optical coherence tomography screening were subjected to visual function assays using optokinetic (OKN) testing and superior colliculus (SC) electrophysiology. At approximately 25 weeks of age (21 weeks after surgery), the eyes were examined histologically for cell survival, phagocytosis, and local toxicity.
Eighty-seven percent of the rCPCB-RPE1-implanted animals showed hESC-RPE survivability. Significant numbers of outer nuclear layer cells were rescued in both group 1 (rMSPM+VN) and group 2 (rCPCB-RPE1) animals. A significantly higher ratio of rod photoreceptor cells to cone photoreceptor cells was found in the rCPCB-RPE1-implanted group. Animals with rCPCB-RPE1 implant showed hESC-RPE cells containing rhodopsin-positive particles in immunohistochemistry, suggesting phagocytic function. Superior colliculus mapping data demonstrated that a significantly higher number of SC sites responded to light stimulus at a lower luminance threshold level in the rCPCB-RPE1-implanted group. Optokinetic data suggested both implantation groups showed improved visual acuity.
These results demonstrate the safety, survival, and functionality of the hESC-RPE monolayer transplantation in an RPE dysfunction rat model.
确定接种于聚合物基质（rCPCB-RPE1植入物）上并植入皇家外科学院（RCS）大鼠视网膜下（SR）间隙的人胚胎干细胞衍生的视网膜色素上皮（hESC-RPE）细胞的安全性、存活率和功能。
将在聚对二甲苯膜上培养的hESC-RPE细胞单层移植到4周龄RCS大鼠的SR间隙。第1组（n = 46）接受无细胞的玻连蛋白包被的聚对二甲苯膜（rMSPM+VN），第2组（n = 59）接受rCPCB-RPE1植入物，第3组（n = 13）作为对照组。基于光学相干断层扫描筛选选择的动物使用视动（OKN）测试和上丘（SC）电生理学进行视觉功能测定。在大约25周龄（手术后21周）时，对眼睛进行组织学检查以评估细胞存活、吞噬作用和局部毒性。
87%植入rCPCB-RPE1的动物显示hESC-RPE细胞存活。第1组（rMSPM+VN）和第2组（rCPCB-RPE1）动物的大量外核层细胞均得到挽救。在植入rCPCB-RPE1的组中发现视杆光感受器细胞与视锥光感受器细胞的比例显著更高。植入rCPCB-RPE1的动物在免疫组织化学中显示hESC-RPE细胞含有视紫红质阳性颗粒，提示具有吞噬功能。上丘图谱数据表明，在植入rCPCB-RPE1的组中，在较低亮度阈值水平下，有显著更多的SC位点对光刺激有反应。视动数据表明两个植入组的视力均有所改善。
这些结果证明了hESC-RPE单层移植在RPE功能障碍大鼠模型中的安全性、存活率和功能。
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