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A scalable retinal differentiation system for the production of substantially mature mini retinas for modeling human retinal development and inherited retinal degenerations

About the Solution

Here we present our solution to the 3-D ROC. Retinal structures from hESCs or hiPSCs have been generated using a number of protocols, but the requirement of manual manipulation, low efficiency, immaturity and variability limit their applications. We have established a retinal differentiation protocol for generating and isolating large quantities of retinal organoids that produce stratified mini retinas from hESCs. The novelties in our solution are the ease of use, scalability, robustness, reproducibility, and the maturity of photoreceptors. The advantages of our protocol are the efficient generation of early retinal epithelium through Matrigel-induced cyst formation and scalable isolation of self-organized retinal organoids through Dispase-mediated cell detachment and subsequent floating culture. Our retinal organoids produce stratified neuroretinal tissues with all five neuronal retina cell types. Notably, outer segments of photoreceptors and outer limiting membrane are evidenced by immunostaining and electron microscopy. We are using the retinal organoids to model optic cup invagination and Leber Congenital Amaurosis. We propose solutions to faithfully recapitulate the complexity of the retina through tissue engineering.

 

 

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