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Preliminary observations on human trabecular meshwork cells in vitro
Abstract:
This report presents our preliminary observations on the trabecular meshwork from human eyes up to 5 days post-mortem in tissue culture. Satisfactory primary cultures were obtained from about 20% of the 423 explants which were investigated. The period prior to growth was from 4 days to 4 weeks and from the appearance of the initial outgrowth it took 25 to 30 days to reach maximum cellular spread within the culture chambers. The progress of the explant and the spreading of the trabecular meshwork cells was monitored by phase-contrast microscopy, time-lapse cinephotomicrography, light microscopy, transmission electron microscopy, scanning electron microscopy and autoradiography (using tritiated thymidine). On the basis of their ultrastructural appearance the cultured meshwork cells seems to be metabolically active. Their cytoplasm contained abundant rough endoplasmic reticulum, many mitochondria, a well developed Golgi apparatus and many coated and uncoated micropinosomes. However, even in short-term culture the trabecular meshwork cells had adapted to the artificial environment of our system and no longer resembled "normal" trabecular meshwork cells as seen in vivo. Since trabecular meshwork cells can quickly adapt their morphology in a culture environment and because the adult human meshwork contains a significant population of non-trabecular cells, the value of long term culture as a means of investigating the cellular activity of the normal and glaucomatous outflow system must be open to question.
Insights
Human trabecular meshwork cells in tissue culture showed metabolic activity but adapted quickly, altering their morphology. This rapid adaptation raises questions about the value of long-term cultures for studying glaucoma.
Area of Science:
- Ophthalmology
- Cell Biology
- Tissue Engineering
Background:
- The trabecular meshwork (TM) is crucial for regulating intraocular pressure.
- Understanding TM cell behavior in vitro is vital for glaucoma research.
- Previous studies have explored TM cell cultures with varying success.
Purpose of the Study:
- To observe the behavior and morphology of human trabecular meshwork cells in short-term tissue culture.
- To assess the metabolic activity and ultrastructural characteristics of cultured TM cells.
- To evaluate the suitability of long-term TM cell cultures for investigating the glaucomatous outflow system.
Main Methods:
- Human donor eyes (up to 5 days post-mortem) were used for explant cultures.
- Phase-contrast microscopy, time-lapse cinephotomicrography, and electron microscopy were employed.
- Autoradiography with tritiated thymidine was used to assess cellular activity.
- Ultrastructural analysis examined cytoplasmic features like endoplasmic reticulum and mitochondria.
Main Results:
- Satisfactory primary cultures were achieved from approximately 20% of explants.
- Cultured TM cells exhibited metabolic activity, with abundant organelles.
- Significant morphological adaptation to the culture environment was observed within short-term cultures.
- Cultured cells no longer resembled in vivo TM cells.
Conclusions:
- Human trabecular meshwork cells are metabolically active in vitro.
- Rapid morphological adaptation of TM cells in culture limits their resemblance to in vivo cells.
- The value of long-term TM cell cultures for studying normal and glaucomatous outflow systems is questionable due to adaptation and the presence of non-TM cells.