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Kapil Bharti, Janine Davis, Juliet Hartford, Vladimir Khristov, Qin Wan, Mostafa Reza Lotfi, Kiyoharu j Miyagishima, Arvydas Maminishkis, Juan Amaral, Sheldon S Miller; Engineering iPS Cell Derived RPE Tissue Using Biodegradable Scaffolds. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5245.
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The recent preliminary success with embryonic stem (ES) cell derived retinal pigment epithelium (RPE) delivered in suspension has provided optimism for a treatment for degenerative eye diseases. Induced pluripotent stem (iPS) cells provide an alternate and an autologous source of stem cells with fewer immune-challenges as compared to ES cells. RPE sheets allow delivery of polarized cells that function more efficiently as compared to cell suspension and can provide an efficacious cell-based product. Here, we aim to develop an autologous iPS cell derived RPE tissue sheet using biodegradable scaffolds.
RPE cells derived from iPS cells using a directed differentiation protocol were used to derive mature, fully-polarized RPE sheets on PLGA and Collagen scaffolds. RPE sheets were authenticated using gene expression, immunofluorescence, electron microscopy, vectorial fluid transport, and measurements of membrane properties of RPE and intracellular calcium measurements.
Fully-polarized, and pigmented monolayer of RPE cells can be grown on PLGA scaffolds. Electron microscopy and immunostaining confirmed the presence of typical RPE characteristics such as apical processes, apically localized pigment granules, tight junctions, and basal infoldings. Electrophysiology and physiology experiments confirmed the polarized nature of cells, the presence of functional channels on the apical and basal sides on the cells and the ability to activate intracellular signaling pathways.
We have engineered and fully-authenticated an RPE tissue using iPS cell derived RPE cells and biodegradable PLGA scaffold. This tissue strongly resembles the native RPE tissue in its morphological and physiological properties. Currently, we are testing this tissue in pre-clinical animal models and developing Good Manufacturing Practices for clinical-grade manufacturing of this tissue.
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