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Xiufeng Zhong, Shengxu Liu, Bingbing Xie, xiaojing song, Dandan Zheng, Liwen HE, Guanjie Gao; Enrichment and characterization of self-forming RPE Spheroids concurrently generated with Retinal Organoids from hiPSCs. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3323.
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© ARVO (1962-2015); The Authors (2016-present)
To enrich and expand hiPSC-derived retinal pigment epithelium (RPE) concurrently generated with neural retina on retinal organoid induction platform
hiPSCs were cultured on MatriGel coated plates with mTeSR1 medium, and differentiated into RPE cells with a published retinal organoid induction method. After detachment of neural retina on 4th wk, the remaining mixture was also scraped off from the dish and subjected to suspension culture for the formation of RPE spheroids. RPE cells were isolated for expansion. Different assays were used to evaluate the biological charateristics of RPE cells.
Under suspension culture, hiPSC-RPE spheroids with pigmentation self-formed, and were readily enriched by removing out the non-retinal tissues. RPE cells could be readily purified from the spheroids, cultured and serially passaged in serum medium, yielding large numbers of cells with high quality in a short period. When switched to serum-free medium, the passaged RPE cells could mature in cellular, molecular and physiological levels.
A simple and novel RPE spheroids formation approach was established to enrich and expand hiPSC-RPE cells generated along with retinal neurons on a universal retinal organoid induction platform. This achievement will evidently reduce the cost and time in producing retinal cells for basic and translational researches, for retinal cell therapy in particular.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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