(770f) Dynamic Culture of Trabecular Meshwork Cells in 3D Biomimetic Scaffolds

Authors: 
Osmond, M., Colorado School of Mines
Dynamic Culture of Trabecular Meshwork Cells in 3D Biomimetic Scaffolds

Matthew Osmond1, Mina B. Pantcheva2, and Melissa D. Krebs1

1 Chemical & Biological Engineering, Colorado School of Mines, Golden, CO

2 Ophthalmology, University of Colorado School of Medicine, Aurora, CO

mina.pantcheva@ucdenver.edu and mdkrebs@mines.edu

Glaucoma is a disease in which damage to the optic nerve leads to progressive, irreversible vision loss. The intraocular pressure (IOP) is the only modifiable risk factor for glaucoma. Strategies for decreasing IOP are useful in preventing or slowing down the progression of glaucomatous neuropathy. Elevated intraocular pressure associated with glaucoma is due to increased aqueous humor outflow resistance, primarily through the trabecular meshwork (TM) of the eye. Currently researchers trying to screen new glaucoma therapeutics or investigate TM cell behavior under certain conditions use TM cells cultured on 2D plastic substrates, a poor model for the actual in vivo tissue. To better simulate the TM tissue, we have created biomimetic collagen - glycosaminoglycan (GAG) scaffolds with structured architecture on which TM cells can be cultured. Using freeze casting techniques, the pore size and pore alignment can be varied to increase cell viability and proliferation. We have demonstrated that TM cells can be successfully cultured on collagen-GAG scaffolds under static culture conditions, with cells proliferating and viable for up to two weeks. These cells have been shown to express myocilin, a marker of TM cells, when exposed to dexamethasone just as they do when cultured on 2D cell culture dishes. Now we are investigating the influence of various pore size, pore alignment and chemistry of the surrounding matrix on these TM cells under both static and dynamic (perfusion) culture conditions. With our work, we aim to improve the understanding of TM cell biology, and to aid in the screening and development of new therapeutic agents for glaucoma.

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