Simplified and reproducible two-step differentiation method optimized for efficient differentiation of human-induced pluripotent stem cells into retinal progenitor cells and functional retinal ganglion cells   

More than three million Americans live with glaucoma – a disease that can cause blindness resulting from optic nerve damage. In glaucoma, retinal ganglion cells (RGCs) - a type of nerve cell found in the retina that processes visual information from the eye to the brain - are killed and cannot be naturally replaced. Therefore, transplantation of RGCs into the eye to replace dead RGCs promises to rescue vision loss in glaucoma patients and patients with RGC loss due to traumatic brain injury. Human-induced pluripotent stem cells (hiPSCs) can be differentiated into functional RGCs, yet differentiation methods remain limited due to their complexity, irreproducibility, low yield, and use of genomic manipulation. 

A simplified, two-step human iPSC differentiation method optimized for the effective and reproducible generation of functionally mature RGCs. The methodology is highly efficient, enabling a highly scalable process for RGC generation. Moreover, it solely employs chemical agents for differentiation and avoids safety concerns involving genetic manipulation.   

• Use of chemically-defined medium avoids off-target risks involving genetic modification
• High yield of RGCs (>95% of cells)
• Simultaneous production of retinal progenitor cells which can be used for transplantation to rescue photoreceptor loss
• Does not require cell sorting for enrichment of RGC population in culture
• Simplified technique in 2D monolayer avoids complexity and variability involving 3D organoid use
• iPSC-derived RGCs are mature and electrophysiologically functional (i.e. can generate action potentials)
• iPSC-derived RGCs when transplanted into mouse eyes integrate and extend axons through optic nerve head