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Carla Bernadette Mellough, Joseph F Collin, Mahmoud Khazim, Evelyne Sernagor, Nicholas Wride, David Steel, Majlinda Lako; The formation of primitive ocular structures and stratified neural retina from human pluripotent stem cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1358.
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Diseases of the outer retina are a leading cause of irreversible blindness worldwide, yet no treatments currently exist for many forms. The ability to generate retinal tissue for disease modelling, drug screening or transplantation would be extremely useful in order to resolve this important challenge. Human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC) can be differentiated in vitro to develop towards retinal cells including photoreceptors and retinal pigmented epithelium (RPE). Their ability to self-organise into complex retinal tissue, similar to embryonic human retina, has recently been demonstrated and represents a leap towards in vitro modelling of disease phenotypes and the study of human retinal ontogenesis. The reproducibility and efficacy of this capability is however not well established across multiple hESC/hiPSC lines. We have built upon our previous work to devise a simple and reliable method that produces laminated retinal tissue in vitro using one factor only.
hESC/hiPSC lines were expanded on mouse embryonic fibroblasts then differentiated under 3-dimensional conditions for 90 days in a ventral neural induction medium either alone or supplemented with human insulin-like growth factor (IGF).
The addition of IGF during differentiation orchestrates the formation of ocular-like structures from hESC/hiPSC with high frequency compared to controls. IGF-generated structures contain not only RPE and neural retina, but other eye elements including primitive lens and corneal epithelium. Retinal organisation is reminiscent of developing human retina and comprises multiple phenotypes including photoreceptors, amacrine and ganglion cells, which establish synaptic connections and form visible plexiform layers. Photoreceptors exhibit primitive rod- and cone-like inner and outer segments and functional cyclic nucleotide gated channel properties. Inhibition of IGF pathway signalling abolished the ability of hESC/hiPSC to give rise to viable laminated retina, establishing an important role for this pathway in retinal histogenesis.
The efficient derivation of ocular derivatives including laminated retinal tissue can be achieved from hESC and hiPSC using IGF, offering exciting new opportunities to study retinal development and disease and to provide an expandable source of transplantable derivatives for cell replacement studies.
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