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Joo Young Shin, Hyeong Gon Yu; Simplified and Efficient Generation of Photoreceptors from Human Pleuripotent Stem Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4050.
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© ARVO (1962-2015); The Authors (2016-present)
Despite numerous attempts to generate photoreceptors from human pleuripotent stem cells, the problem of efficiency and long periods of time required for photoreceptor derivation remain one of the many obstacles in the introduction of these cells in the therapy of retinal degenerative diseases. We report our simplified and efficient differentiation method into photoreceptors using our previously reported spherical neural mass derived from human embryonic stem cells.
Spherical neural masses were broken down and replated and co-cultured with retinal pigment epithelial cells such as ARPE-19 or human embryonic stem cell-derived retinal pigment epithelial cells, in neural differentiation medium with N2 and B27.
Spherical neural masses are neural progenitor cell masses derived from human embryonic stem cells, and are cryo-preservable for extended periods of time. When replated and co-cultured with retinal pigment epithelial cells, differentiation towards photoreceptors could be detected within 10-14 days in minimal culture conditions. These cells formed long neurites and expressed markers specific for rod and cone photoreceptor cells such as RHODOPSIN, OPN1SW, and OPN1LM, identified by polymerase chain reaction and fluorescence immunocytochemistry. These cells also expressed components of the phototransduction cascade such as transducin, rhodopsin, peripherin, and Pde6b.
Generation of photoreceptors from human embryonic stem cell-derived spherical neural mass with a simplified and efficient method using retinal pigment epithelial cell co-culture with minimal addition of supplements may faciliate the use of these cells in the therapy of retinal degenerative diseases.
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