Markers and antibodies for characterization of goat mammary tissue and the derived primary epithelial cell cultures

Abstract

A selection of commercially available antibodies, targeted against markers employed in studies of mammary gland biology, was tested to determine their reactivity in goat mammary tissue and the derived tissue cultures. Expression of the markers smooth muscle actin (SMA), selected keratins (KRT) 5, 14, 18, and 19, CD24 molecule (CD24), epithelial cell adhesion molecule (EPCAM), mucin 1 (MUC1), integrin subunit alpha 6 (ITGA6; CD49F), integrin subunit beta 1 (ITGB1; CD29), cyclin dependent kinase inhibitor 1A (CDKN1A; p21), membrane metalloendopeptidase (MME; CD10), progesterone receptor (PGR), estrogen receptor 1 (ESR1), and vimentin (VIM) was first assessed on mRNA level, using reverse transcription PCR (RT-PCR). The reactivity of the antibodies in the tissue sections and the derived tissue cultures was determined using immunofluorescence. The result of this study is a list of commercially available antibodies, raised mostly against human antigens, which also recognize orthologous goat antigens and are useful for characterization of different mammary cell types. Additionally, primers that are functional in detecting expression of mammary lineage markers in goat mammary mRNA isolates were validated. The suggested antibodies, PCR primers, and the described methods are of practical value for researchers interested in characterization and isolation of cell types comprising mammary tissue of goats and probably other ruminants