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The effect of organ culture on human trabecular meshwork
1Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota 55905.
Abstract:
The effect of organ culture on the trabecular meshwork was studied in a series of human eyes using a perfusion culture system. One eye of a pair was cultured while the fellow eye was immediately fixed in glutaraldehyde. Culture periods ranged from 2 to 28 days, during which a mean intraocular pressure of 27 mmHg +/- 15 was maintained. The perfused culture medium appeared to leave the eye via Schlemm's canal and collector channels, mimicking the in vivo situation. The trabecular meshwork was well maintained with this system in 21 of 25 eyes. Cells remained in position on trabecular lamellae, cellular organelles usually remained normal, and the lamellae remained intact. Culture-induced changes were noted, with some cells developing intracytoplasmic lipid vacuoles and other cells developing swollen mitochondria. In addition, scattered focal cell necrosis was observed, most often in the uveal meshwork. Meshwork cellularity (nuclei:solid tissue) was determined with an image analysis system; there was an average cell loss of 20-40% in the cultured eyes as compared to their respective fellow control eyes. Overall, organ culture appears to maintain the human trabecular meshwork for at least 28 days, allowing controlled experimental studies to be performed.
Insights
Human trabecular meshwork organ culture using a perfusion system effectively maintained tissue structure for up to 28 days. This method supports controlled experimental studies of the trabecular meshwork.
Area of Science:
- Ophthalmology
- Cell Biology
- Tissue Engineering
Background:
- The trabecular meshwork is crucial for regulating intraocular pressure.
- Understanding its behavior in vitro is essential for glaucoma research.
- Previous methods for culturing ocular tissues have limitations.
Purpose of the Study:
- To evaluate the efficacy of a human eye organ culture system using perfusion.
- To assess the structural and cellular integrity of the trabecular meshwork during culture.
- To determine the suitability of this system for experimental manipulation.
Main Methods:
- Human donor eyes were used, with one eye undergoing perfusion organ culture and the fellow eye serving as a control.
- A mean intraocular pressure of 27 mmHg was maintained throughout culture periods ranging from 2 to 28 days.
- Trabecular meshwork cellularity was quantified using image analysis.
Main Results:
- The perfusion system successfully mimicked in vivo aqueous humor outflow pathways.
- Trabecular meshwork structure was well-preserved in 21 out of 25 cultured eyes.
- Minor culture-induced changes included lipid vacuoles and swollen mitochondria, with a 20-40% cell loss observed.
Conclusions:
- Human organ-cultured trabecular meshwork remains viable and structurally sound for at least 28 days.
- This perfusion culture system provides a reliable model for studying trabecular meshwork physiology and pathology.
- The system facilitates controlled experimental investigations relevant to intraocular pressure regulation and glaucoma.