Protocols for TM Cell Replacement Using Differentiated TM-Like Induced Pluripotent Stem Cells (iPSC)

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Induced pluripotent stem cells are differentiated into TM-like iPSCs and transplanted into a cell depletion glaucomatous cell loss model. This restores the capability of this cell loss-perfused human anterior segment model to respond to a 2X pressure challenge with a normal IOP homeostatic response. Glaucomatous anterior segments cannot adequately respond to a 2x pressure challenge. Herein, we present protocols for working with human iPSCs, differentiating iPSCs into TM-like iPSCs, establishing the saponin TM cell depletion perfusion model for cell loss in the anterior segment, testing IOP homeostatic response, transplanting TM-like iPSCs into perfused human anterior segments, and evaluating transplanted cell integration. These protocols should facilitate similar studies on glaucomatous anterior segments and translation of these approaches to treat glaucomatous TM cell loss and their compromised IOP homeostatic response.