Generating cell-derived matrices from human trabecular meshwork cell cultures for mechanistic studies

Area of Science:

• Ophthalmology
• Biomaterials Science
• Cell Biology

Background:

• Ocular hypertension is linked to trabecular meshwork (TM) extracellular matrix (ECM) changes, impacting aqueous humor outflow.
• Existing in vivo and ex vivo models have limitations in dissecting complex molecular mechanisms.
• In vitro models manipulating single ECM properties lack multifactorial relevance.

Purpose of the Study:

• To present a streamlined protocol for generating cell-derived matrices (CDMs) from TM cells.
• To demonstrate methods for pharmacologically manipulating TM cells and CDMs (e.g., glucocorticoid-induced CDMs, genipin treatment).
• To highlight the utility of CDMs for mechanistic studies of TM ECM and potential applications in regenerative medicine.

Main Methods:

• Generation of CDMs from normal and glaucomatous donor TM cell cultures.
• Pharmacological manipulation of TM cells to induce specific CDM properties (glucocorticoid treatment).
• Chemical modification of CDMs using reagents like genipin.

Main Results:

• A streamlined protocol for CDM generation was established.
• Methods for creating pharmacologically and chemically modified CDMs were documented.
• CDMs provide a tunable 3D microenvironment mimicking native ECM characteristics.

Conclusions:

• CDMs offer a superior in vitro model for studying TM ECM in ocular hypertension due to their 3D, multifactorial nature.
• These matrices can be tailored to mimic pathological conditions and facilitate mechanistic investigations.
• CDMs hold promise for future research in TM regenerative strategies.