Development of a fibroblast proliferation bioassay to detect mediators of pulmonary fibrosis.
Reist-R; Bryner-K; Wearden-P; Blackford-J; Vrana-K; Castranova-V; Dey-R
Proceedings of the 3rd Symposium on Respirable Dust in the Mineral Industries, Pittsburgh, Pennsylvania, October 17-19, 1990. Frantz RL, Ramani, RV, eds., Littleton, CO: Society for Mining, Metallurgy, and Exploration, 1990 Oct; :179-181
The effects of platelet derived growth factor (PDGF) and 2% rat serum, as a source of competence factor, on mitogenesis was investigated. Sprague-Dawley-rat lung fibroblasts were isolated and human fetal lung fibroblasts obtained from the American Tissue Culture Collection (ATCC153). After plating, cells were rested for 48 hours (hr) in plasma free media, then exposed to 150 nanograms/milliliter (ng/ml) PDGF and 2% rat plasma, separately or in combination for 6hr. Incorporation of tritiated thymidine during a 24hr labeling period in plasma free medium following the test treatments provided a measurement of mitogenesis. Positive controls were 10% and 20% fetal calf serum for rat and human fibroblasts, respectively. Serum free medium served as the negative control. Significant mitogenesis did not occur in cells treated with PDGF or 2% rat serum alone. Cells treated with a combination of PDGF and 2% rat serum incorporated significantly more thymidine than the positive controls, indicating activation of mitogenesis. A dose response curve demonstrated the sensitivity of the competence stimulating activity assay to a lower limit of 25ng/ml PDGF. The authors conclude that the described assay for fibrogenic factor production represents an improvement over previously published methods.
Lung-cells; Lung-fibrosis; Growth-factors; In-vitro-study; Mitosis; Respirable-dust; Quantitative-analysis; Bioassays; Pulmonary-disorders; Respiratory-system-disorders; Mammalian-cells
Proceedings of the 3rd Symposium on Respirable Dust in the Mineral Industries, Pittsburgh, Pennsylvania, October 17-19, 1990