title: Evaluation of a replacement method for mammary gland biopsies by comparing gene expression in udder tissue and mammary epithelial cells isolated from milk.
abstract: Somatic cells isolated from milk offer an attractive non-invasive replacement of invasive udder biopsies for monitoring bovine mammary gland metabolism. However, for metabolic gene expression studies the mammary gland epithelial cells (MEC) isolated from milk have to be purified from the non-epithelial leukocyte fraction in milk samples. In our study, enrichment of MEC by using anti-cytokeratin peptide 18 (KRT18) antibody coated magnetic beads was evaluated. MEC showed a substantially increased expression of the epithelial-cell-specific KRT18 gene compared to udder tissue. The expression levels of genes specific for mammary gland epithelial cells (CSN3 and LALBA) showed a significant positive correlation in MEC and also in udder tissue. However, no significant correlation of the expression of a specific gene was found between udder and MEC samples. Therefore, MEC isolated from total milk samples via KRT18 antibodies probably do not reflect the true metabolic situation of the bovine udder. Thus, quantitative gene expression profiling of MEC isolated via KRT18 antibodies has to be interpreted carefully with respect to the situation in the udder.
authors.affiliation: Res. Unit Molecular Biology, Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Krappmann
authors.forename: R
authors.initials: R
authors.lastname: Weikard
authors.forename: C
authors.initials: C
authors.lastname: Kühn
mesh_terms: D000818:Animals; D001706:Biopsy / Q000662:veterinary*; D002417:Cattle; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D007774:Lactation / Q000502:physiology; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000378:metabolism*; D008892:Milk / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012333:RNA, Messenger