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In vitro Culture alters the Expression of Xenobiotic Transporters by Mammary Epithelial Cells
Author(s) -
Gehring Ronette,
Malreddy Pradeep,
Silver Kristopher,
Wang Florence,
Schultz Bruce
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1047.11
Subject(s) - in vitro , xenobiotic , cell culture , messenger rna , epithelium , biology , mammary gland , microbiology and biotechnology , gene expression , abcg2 , transporter , atp binding cassette transporter , biochemistry , gene , enzyme , genetics , cancer , breast cancer
Some drugs and toxins are actively transported across the mammary epithelial barrier, resulting in high milk concentrations. To assess potential exposure through milk, in vitro models of the mammary epithelial barrier are needed. The goal of these studies was to compare the expression of xenobiotic transporters between native tissues, cultures of freshly isolated bovine mammary epithelial cells and a bovine mammary epithelial cell line (BME‐UV). Real time PCR was employed to evaluate the expression of mRNA coding for 5 xenobiotic transporters in mammary tissue isolated from lactating dairy cows, primary epithelial cells isolated from these tissues and cultured in T25 flasks or as polarized monolayers on permeable supports, and BME‐UV cells cultured as polarized monolayers on permeable supports. 18S rRNA was employed as an internal standard for comparison. Relative to the native tissue, the expression of mRNA coding for ABCB1 and NCBT1 were increased in all the cell cultures, whereas ABCG2 and OATPD mRNA expression were reduced. The expression of OCT1 mRNA remained unchanged. These initial results suggest that standard culture conditions promote the expression of putative drug transport proteins in proportions and levels that do not directly reflect those observed in the actively lactating dam. Further studies are needed to refine culture conditions for these potential in vitro models. [NIH P20‐ RR017686 Core C]