Mastitis, inflammation of the mammary gland, is one of the most economically important diseases of dairy ruminants. Mastitis is usually caused by bacteria. Cell cultures can serve as an alternative to live animal studies and provide an in vitro model to study the course of the disease, which is crucial for the establishment of successful mastitis prevention strategies. In the experimental part of the thesis, we established a primary cell culture of bovine mammary epithelial cells – pbMEC, and used it as a cell model to study mastitis. pbMEC were established in a non-invasive manner from milk, and the presence of epithelial cells was confirmed by immunofluorescence. The cells were immunostimulated with the Gram negative bacterial virulence factor LPS and incubated for 3 and 24 hours. After the incubation, RNA was isolated, reverse transcribed into cDNA and the differential expression of the IL-8 and TNF-alpha cytokines was determined by qPCR. The results showed that the expression of IL-8 and TNF-alpha increased after immunostimulation with LPS, suggesting that the pbMEC culture is immunocompetent. The expression of IL-8 was higher after 24 hours than after 3 hours, while the expression of TNF-alpha was higher after 3 hours and was only slightly elevated after 24 hours. The results are consistent with the findings of previous studies and suggest that pbMEC culture is a potentially useful in vitro model to study the immune response during mastitis.
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