Human trabecular meshwork organ culture

Human trabecular meshwork obtained from postmortem eyes was maintained in organ culture condition up to 12 weeks. The explants were dissected into specimens of three different sizes. Types 1 and 2 specimens were larger than 4 mm square and 1 mm thick, and type 3 specimens were about 1 mm3. Each specimen was placed in a depression made in the center of an agar block and the block was placed in a plastic culture dish. The space between the agar block and the culture dish was filled with liquid medium consisting of MEM Eagle supplemented with 10% fetal bovine serum, 10% chick embryo extract and 200 U/ml penicillin G. Afterwards, the explant was incubated in 5% CO2 with 100% humidity. Types 1 and 2 specimens maintained a normal appearance on electron microscopy during the entire 12 weeks of culture, while type 3 specimens showed marked cellular activation and cell loss soon after explantation. Morphometric studies regarding cell number in sections for light microscopy obtained from types 1 and 2 specimens revealed good preservation of trabecular cells during the 12 weeks of culture. Our culture system may be useful in investigating the direct effects produced on human trabecular meshwork by photocoagulation and various agents, for example anti-glaucoma drugs, steroids and viscoelastic materials.