Generation of highly enriched populations of optic vesicle-like retinal cells from human pluripotent stem cells.
Generation of highly enriched populations of optic vesicle-like retinal cells from human pluripotent stem cells.
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DOI：
10.1002/9780470151808.sc01h08s32
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发表时间：
2015-02-02
影响因子：
--
通讯作者：
Meyer, Jason S
中科院分区：
文献类型：
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作者：
Ohlemacher, Sarah K;Iglesias, Clara L;Sridhar, Akshayalakshmi;Gamm, David M;Meyer, Jason S
下文描述了从人多能干细胞（hPSC）有效且可再现地分化视网膜细胞的程序。通过逐步方案获取细胞，以通过用神经诱导培养基（NIM）处理将它们导向神经命运，然后通过暴露于视网膜分化培养基（RDM）导向视网膜命运。将未分化的hPSC从基质胶包被的板酶促提升并暴露于悬浮液中的NIM。在悬浮液中的分化允许细胞形成三维聚集体。在分化7天时，将聚集体铺板并附着于6孔板，其中开始建立神经上皮命运。在分化16天后，神经球被提升并保持在RDM中以产生三维视泡样结构。该过程允许有效和及时地产生各种视网膜细胞类型，包括神经节细胞、视网膜色素上皮以及视锥和视杆光感受器。使用该方案来产生无数的视网膜细胞类型有利于人类视网膜发生的体外研究，并且将使视网膜功能障碍能够更容易地在体外研究，以及提供大量细胞群体，以帮助药物开发和患者特异性治疗。
The procedure to efficiently and reproducibly differentiate retinal cells from human pluripotent stem cells (hPSCs) is described below. Cells are taken through a stepwise protocol to direct them toward a neural fate by treatment with neural induction medium (NIM), then to a retinal fate by exposure to retinal differentiation medium (RDM). Undifferentiated hPSCs are enzymatically lifted from matrigel-coated plates and exposed to NIM in suspension. Differentiation in suspension allows the cells to form 3 dimensional aggregates. At 7 days of differentiation, aggregates are plated and attach to 6 well plates, where a neuroepithelial fate begins to be established. Upon 16 days of differentiation, neurospheres are lifted and maintained in RDM to create a three-dimensional optic vesicle-like structure. This procedure allows for the efficient and timely generation of a variety of retinal cell types, including ganglion cells, retinal pigment epithelium, as well as cone and rod photoreceptors. The use of this protocol to generate a myriad of retinal cell types facilitates in vitro studies of human retinogenesis, and will enable retinal dysfunction to be more easily studied in vitro, as well as providing a large population of cells with which to aid in drug development and patient specific therapies.