比較研究胎牛血清和馬血清對初級馬支氣管成纖維細(xì)胞的生長和分化的影響

Cell yield from digested bronchial tissue was consistent. Under both culture conditions, EBF were stained with trypan blue and the percentage of viable cells was similar, usually?>?95%. No significant evidence of cell necrosis or cell apoptosis was observed under inverted light microscopic analysis. With regard to cell morphology, EBF cultured in DMEM with 10% FBS appeared to be typically flattened and spindle-shaped (with a homogenous cytoplasm) over several weeks of passages compared to EBF cultured in DMEM with 10% HS (Figure 1A). In medium containing FBS, EBF were grown over the growth surface with only loose cell-cell-contact until reaching confluence and then formed tight parallel lines which remained as typical fibroblastic monolayer until passage 15. EBF between passages 16 – 20 started to change their morphology: cells were large, flat and more polygonal shaped, with a large, heterogeneous nucleus. At the same time, cell growth was rapidly reduced and cell viability was diminished. In contrast, EBF cultured in the presence of HS showed altered morphological changes within 2 days of culture; cells were small and more compact in shape combined with granula-like dark structures in the cytoplasm (Figure 1C). Moreover, cells grew in clusters and chains and failed to reach confluence within a week. This morphological behaviour of EBF was seen under this culture conditions until passage 7, but thereafter, EBF failed to proliferate regularly and decreased in their viability (passage 9). Under both culture conditions, most EBF (>99%) were positive for vimentin, but with more characteristic filamentous structures within the cytoplasm in medium containing FBS (Figure 1B) than in the presence of HS (Figure 1D).

In sum, serum addition to basal EBF medium enhanced EBF differentiation into myofibroblasts, and these findings are useful to develop in vitro fibroblast culture models that mimic in vivo physiological processes and to study airway disease mechanisms and remodeling.

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