Derivation of Primary Human Trabecular Meshwork Cell-Derived Matrices

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Ocular hypertension (OHT) partly results from increased resistance to aqueous humor (AQH) outflow from the anterior chamber of the eye as a result of changes in the extracellular matrix (ECM) of trabecular meshwork (TM). Even though in vivo animal models of OHT often rely upon singular gene knockout systems, these do not reveal the complexities in matrix changes (structure and composition) in the disease. Also, in vitro systems mimic the native topography, mechanics, or biochemistry of native ECM exist but often fail to mimic the multifaceted environment around the cells. There is, thus, a paucity of physiologically relevant assays/models to perform non-clinical mechanism-based research. Cell-derived matrices (CDMs) represent a 3D microenvironment derived from a cell type of interest that partially simulates the matrix similar to conditions in vivo. To mimic homeostasis under normal conditions or pathological states, the CDM content can be altered by the addition of various classes of compounds. A protocol for generating CDMs from isolated and cultured primary TM cells is described in this chapter.