Source: Annual Congress 2002 - Cell biology and genetics of COPD and emphysema
Disease area: Airway diseases

Abstract

In inflammatory lung disorders, red blood cells (RBC) may interact with extracellular matrix (ECM). In previous studies we showed that RBC and RBC conditioned medium (RBC-CM) enhanced lung fibroblast contraction of collagen gels, secretion of IL-8 and fibronectin as well as stimulated neutrophil graulocyte migration in an in vitro transwell system. In the present study we addressed the question whether RBC and RBC-CM had an impact on fibroblast proliferation. Human foetal lung fibroblasts (HFL-1) were cultured in three-dimensional collagen gels, in the presence or absence of purified RBC or RBC-CM (medium was supplemented with 0.5% foetal calf serum (FCS)). The gels were dissolved in collagenase and the cells were analysed for DNA content with Hoechst dye 33258. To further investigate cell proliferation, a colorimetric cell proliferation assay (WST-1) was used. HFL-1 were cultured in 96-well culture plates in the presence or absence of RBC or RBC-CM with 0.5% FCS. Proliferation was measured in a spectrophotometer at 450 nm. DNA analysis showed that there were significantly lower DNA contents in co-culture with RBC or RBC-CM than in cultures with only HFL-1 (82.6±]10.6 (mean±]SD of OD values), P<0.001; 103.3±]12.1 P<0.05 vs. 123.5±]6.7). WST-1 analysis showed that the fibroblast proliferation were significantly decreased in co-culture with RBC compared to the control (384±]36 (mean±]SD of OD-values) vs. 595±]49; P<0.001) whereas there was no significant difference detected between the control and HFL-1 in culture with RBC-CM. Our results indicate that RBC, when interacting with ECM, may participate in altering the proliferation process of the fibroblasts. This might be an important mechanism regulating tissue repair after injury.

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K. Fredriksson, J. Lundahl, S. I. Rennard, M. C. Skold (Stockholm, Sweden; Omaha, United States Of America). Red blood cells inhibit fibroblast proliferation in vitro. Eur Respir J 2002; 20: Suppl. 38, 645

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