Abstract
Mammary cell cultures have been used to evaluate gene expression under many types of treatments. Additionally, culturing explants of mammary gland of lactating animals could allow the evaluation of the effects of bioactive molecules, considering that the tissue maintains its normal cellular activity as it does in the living animal. The objectives of this study were to evaluate mammary gland explants of lactating ewes as an in vitro model to test the gene expression of transcription factors and lipogenic genes involved in milk synthesis. Explants were cultured for 3 or 24 h and subjected or not to a challenge dose of conjugated linoleic acid (CLA = 290 μM of trans-10, cis-12) and; No CLA = culture medium. The Comet Assay was performed to evaluate DNA fragmentation. RNA was extracted, cDNA was synthesized, and RT-qPCR was carried out. The Comet Assay showed absence of DNA damage in culture at 24 h. Collectively, our results showed gene expression of lipogenic genes and alpha lactalbumin, supporting that mammary gland explants of lactating ewes can be cultured for 24 h for molecular studies.
Published Version
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