期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 128, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/56274
关键词
Developmental Biology; Issue 128; retinal pigment epithelial cells (RPE); retinal pigment epithelium; induced pluripotent stem cells; human embryonic stem cells; differentiation; retinal development; cell culture
资金
- Garland Initiative for Vision
- California Institute for Regenerative Medicine (CIRM) [DR1-01444, CL1-00521, TB1-01177, FA1-00616, TG2-01151]
- Vermont Community Foundation
- Breaux Foundation
- Foundation Fighting Blindness Wynn-Gund Translational Research Acceleration Program
We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing a detailed and thorough protocol is to clearly demonstrate each step and to make this readily available to researchers in the field. This protocol results in a homogenous layer of RPE with minimal or no manual dissection needed. The method presented here has been shown to be effective for induced pluripotent stem cells (iPSC) and human embryonic stem cells. Additionally, we describe methods for cryopreservation of intermediate cell banks that allow long-term storage. RPE generated using this protocol might be useful for iPSC disease-in-a-dish modeling or clinical application.
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