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Wei Liu, Albert Lowe, Raven Harris, Punita Bhansali, Ales Cvekl; Characterization of hESC-derived retinal tissues in long-term cultures. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6067.
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© ARVO (1962-2015); The Authors (2016-present)
This study aims to characterize hESC-derived retinal tissues in long-term cultures.
Undifferentiated H1 hESCs were cultured on Matrigel-coated plates in mTeSR medium and passaged every 5 to 7 days with Dispase or ReLeSR. Retinal differentiation was performed with a modified Matrigel-based method. The hESC-derived retinal tissues were cultured in DMEM/F12 medium supplemented with 10% FBS and B27, and were characterized with immunohistochemistry and electronic microscopy.
The retinal tissues in long-term cultures displayed as a laminar structure with all retinal cell types. The retinal cells were identified by immunohistochemistry with these markers: ISLET1/2 and BRN3 for ganglion cells, Tuj1 for ganglion cells and amacrine cells, Syntaxin for amacrine cells, Calbindin 28k for horizontal cells, PKCalpha for bipolar cells, RCVRN for photoreceptor cells, and Gutamine synthetase and LHX2 for Müller glial cells. The photoreceptor cells highly expressed RHO, L/M-Opsin, and S-Opsin. Importantly, the inner and outer segments of the photoreceptor cells in the retinal tissues were identified by the characteristic ultrastructures in electronic microscopy.
Our retinal differentiation system is capable of generating retinal tissues resembling human retinae and thus could be useful in stem cell-based research and applications for retinal disease.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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