Differentiation of Retinal Organoids from Human Pluripotent Stem Cells.
Differentiation of Retinal Organoids from Human Pluripotent Stem Cells.
复制标题
从人类多能干细胞分化出视网膜器官组织。
DOI：
10.1002/cpsc.95
复制
发表时间：
2019-09-01
影响因子：
--
通讯作者：
Lako, Majlinda
中科院分区：
文献类型：
--
作者：
Chichagova, Valeria;Dorgau, Birthe;Lako, Majlinda
本单元描述了一种从人类多能干细胞（hPSCs）生成包含所有主要视网膜细胞类型且对光有反应的视网膜类器官的实验方案。hPSCs在96孔板中进行分化，以便大规模生产类器官，这些类器官可用于多种应用，包括人类视网膜发育研究、疾病建模和化合物筛选。这种分化方法基于这样的认识：胰岛素样生长因子1信号通路与视黄酸和三碘甲状腺原氨酸一起对视网膜发育很重要。在培养22周后，类器官形成一层厚厚的神经上皮，其中包含光感受器以及双极细胞、水平细胞、无长突细胞、米勒细胞和视网膜神经节细胞。通过形态学观察以及免疫细胞化学检测到的蛋白质定位，可以追踪分化进程。© 2019由约翰威立父子公司出版。
This unit describes a protocol for generating retinal organoids that contain all major retinal cell types and are responsive to light from human pluripotent stem cells (hPSCs). hPSCs are differentiated in 96-well plates to allow large-scale production of organoids that could be used for multiple applications, including study of human retinal development, disease modeling, and compound screening. The differentiation approach is based on the knowledge that insulin-like growth factor 1 signaling together with retinoic acid and triiodothyronine is important for retinal development. After 22 weeks in culture, the organoids form a thick layer of neuroepithelium containing photoreceptors and bipolar, horizontal, amacrine, Muller, and retinal ganglion cells. Differentiation progress can be tracked by morphological observations and protein localization, as detected with immunocytochemistry. © 2019 by John Wiley & Sons, Inc.