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LI ZHANG, Danhong Zhu, Hossein Nazari Khanamiri, Biju Thomas, Mark S Humayun; Expression of phagocytosis-related receptors of subretinal implantated hESC-RPE cells in RCS rats. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1139. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Retinal degeneration, including retinitis pigmentosa and age-related macular degeneration, are leading cause of blinding diseases worldwide. Efficient phagocytosis of photoreceptor outer segment (POS) fragments by retinal pigment epithelial (RPE) cells, which depends on the apical distribution of the integrin family adhesion receptor αvβ5 and Mer tyrosine kinase (MerTK) receptors, plays a key role in biological renewal and maintenance of retina health. hESC-RPE cell based treatment for retinal degeneration is heading towards clinical trial. How does the implanted hESC-RPE cell conduct its biology function and express functional-related receptors, remains currently unknown. This study aimed to observe the expression of functional-related receptors of hESC-RPE after implanted into the subretinal space of retinal degenerated animal model.
Polarized monolayer hESC-RPE cell cultured on an ultrathin scaffold were transplanted into the subretinal space of RCS rats, which has been wildly used as an animal model of retinal degeneration. Eyeball samples were fixed and dissected at different time point (1 week, 2 weeks, 1 month and 4 months) after implantation. Optical coherence tomography (OCT) and H&E staining were used to assess the position of implanted scaffold in retina. Expression of human cell marker (TRA-1-85) and RPE specific marker (RPE65) were observed by Immunofluorescence (IF) staining. Phagocytosis-related receptors, including Integrin αvβ5, MerTK and Rhodopsin were stained with corresponding antibodies.
OCT and H&E staining confirmed the presence of monolayer of hESC-RPE attached to parylene up to 4 months post-implantation. No expression of Integrin αvβ5, MerTK or Rhodopsin was observed by IF staining until 2 weeks after implantation. Long-term expression of Phagocytosis-related receptors was observed 1 month and 4 months after implantation
Subretinal implanted hESC-RPE cell might not conduct its biology function in short-term after implantation. Microenvironment of degenerated retina or surgical trauma might attribute to the delayed expression of functional receptors of hESC-RPE cell.
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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