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Homologous serum-stimulated fibronectin synthesis in human retinal pigmented epithelial cells
S Z Stocks1, I A Shiels, S M Taylor
1Department of Physiology and Pharmacology, University of Queensland, St Lucia, Brisbane, Australia. zhang@plpk.uq.oz.au
Purpose:
Fibronectin expression has been recorded in subretinal membranes from patients with proliferative vitreoretinopathy (PVR). Retinal pigmented epithelial (RPE) cells are a major cellular component of PVR membranes and synthesize fibronectin. We investigated the effects of human serum (as a model of vascular leakage in vivo) on the expression of fibronectin by cultured human RPE cells and compared the responses to those stimulated by fetal bovine serum (FBS).
Methods:
Human RPE cells were incubated in M199 with 1, 10 and 40% human adult serum or FBS for 72 h.
Results:
Retinal pigmented epithelial cells expressed maximum extracellular matrix fibronectin when exposed to 40% human serum using immunohistochemistry. Using ELISA to quantify fibronectin, 10 and 40% human serum significantly increased the total fibronectin (P < 0.01; n = 4), whereas FBS did not affect fibronectin expression.
Conclusions:
The results show that human serum contains substances stimulating fibronectin synthesis in human RPE cells.
Insights
Human serum significantly increases fibronectin expression in retinal pigmented epithelial (RPE) cells, a key component in proliferative vitreoretinopathy (PVR). This suggests human serum contains substances that stimulate fibronectin synthesis in RPE cells.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- Fibronectin is present in subretinal membranes associated with proliferative vitreoretinopathy (PVR).
- Retinal pigmented epithelial (RPE) cells are a primary cellular component of PVR membranes and are known producers of fibronectin.
Purpose of the Study:
- To investigate the impact of human serum, mimicking in vivo vascular leakage, on fibronectin expression by cultured human RPE cells.
- To compare the effects of human serum with fetal bovine serum (FBS) on RPE cell fibronectin synthesis.
Main Methods:
- Human RPE cells were cultured in M199 medium.
- Cells were exposed to varying concentrations (1%, 10%, 40%) of either human adult serum or FBS for 72 hours.
- Fibronectin expression was assessed using immunohistochemistry and ELISA.
Main Results:
- Immunohistochemistry revealed maximal extracellular matrix fibronectin expression in RPE cells treated with 40% human serum.
- ELISA quantification demonstrated that 10% and 40% human serum significantly increased total fibronectin levels (P < 0.01).
- FBS did not significantly alter fibronectin expression in the cultured RPE cells.
Conclusions:
- Human serum contains factors that stimulate fibronectin synthesis in human RPE cells.
- These findings highlight a potential mechanism involving human serum in the pathogenesis of PVR through RPE fibronectin production.