Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Science Key Laboratory.
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Science Key Laboratory;
J Vis Exp. 2021 Apr 21(170). doi: 10.3791/62298.
Retinal cell transplantation is a promising therapeutic approach, which could restore the retinal architecture and stabilize or even improve the visual capabilities to the degenerated retina. Nevertheless, progress in cell replacement therapy presently faces the challenges of requiring an off-the-shelf source of high quality and standardized human retinas. Therefore, an easy and stable protocol is needed for the experiments. Here, we develop an optimized protocol, based on a self-organizing method with the use of exogenous molecules and reagent A as well as manual excision to generate the three-dimensional human retina organoids (RO). The human Pluripotent Stem Cells (PSCs)-derived RO expresses specific markers for photoreceptors. With the addition of COCO, a multifunctional antagonist, the differentiation efficiency of photoreceptor precursors and cones is significantly increased. The efficient use of this system, which has the benefits of cell lines and primary cells, and without the sourcing issues associated with the latter, could produce confluent retinal cells, especially photoreceptors. Thus, the differentiation of PSCs to RO provides an optimal and biorelevant platform for disease modelling, drug screening and cell transplantation.
视网膜细胞移植是一种很有前途的治疗方法，它可以恢复视网膜的结构，并稳定甚至改善退化视网膜的视觉功能。然而，细胞替代疗法的进展目前面临着需要现成的高质量和标准化人类视网膜来源的挑战。因此，需要一种简单稳定的实验方案。在这里，我们开发了一种优化的方案，该方案基于使用外源性分子和试剂 A 以及手动切除的自组织方法来生成三维人视网膜类器官 (RO)。人多能干细胞 (PSC) 衍生的 RO 表达光感受器的特异性标志物。加入多功能拮抗剂 COCO 后，光感受器前体细胞和锥体的分化效率显著提高。该系统的高效利用具有细胞系和原代细胞的优点，而没有后者与来源相关的问题，可以产生大量的视网膜细胞，特别是光感受器。因此，将 PSC 分化为 RO 为疾病建模、药物筛选和细胞移植提供了一个最佳的、有生物学相关性的平台。
