Preparation of Human Trabecular Meshwork Cells for Metabolomic Studies

The trabecular meshwork (TM) is a sieve-like structure within the anterior chamber of the eye responsible for the drainage of aqueous humor and regulation of intraocular pressure (IOP). Dysfunctional TM causes resistance to aqueous outflow and ocular hypertension (elevated IOP), the only modifiable risk factor in the development of glaucoma. The study of metabolites, small bioactive compounds derived from metabolic processes, is a powerful tool for gaining insights into the mechanism of disease and development of drug therapies and biomarkers. Metabolomic studies of primary trabecular meshwork cells can yield insights into the pathophysiology of ocular hypertension as well as identify novel targets for the treatment of glaucoma. This protocol chapter will detail the dissection, culturing, and preparation of human trabecular meshwork cells for metabolite extraction.