Publikationsdatenbank

Establishment of in vitro models of bovine lactocytes and myoepithelial cells (2025)

Art

Poster

Autoren

Rickmann, Kai (WE 1)

Kipar, Claudia (WE 1)

Al Masri, Salah (WE 1)

Nshdejan, Arpenik (WE 1)

Bahramsoltani, Mahtab (WE 1)

Kongress

XXXVth EAVA Congress 2025

Toulouse, 22. – 25.07.2025

Quelle

Sprache

Englisch

Kontakt

Institut für Veterinär-Anatomie

Koserstr. 20
14195 Berlin
+49 30 838 75784
anatomie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

Due to concerns regarding animal welfare and climate protection, there is a particular interest in developing alternative methods of milk production. A suitable in vitro model could represent such an alternative. For this purpose, primary cells were isolated from udder tissue of slaughtered, lactating cows and cultivated in cell culture flasks in a medium containing RPMI 1640 medium, fetal calf serum and penicillin-streptomycin.
During cultivation, three different cell types could be distinguished phenotypically using light microscopy: slender, elongated cells, compact, cobblestone-like cells and individual spindle-shaped cells in between. Using immunocytochemistry, the slender, elongated cells were identified as myoepithelial cells using an antibody against alpha smooth muscle actin, and the spindle-shaped cells were identified as fibroblasts using an antibody against collagen I. The positive controls were conducted immunohistochemically on tissue sections from the bovine udder.
For the establishment of monocultures of luminal, milk-producing lactocytes, myoepithelial cells or co-cultures of both cell types, fluorescence-activated cell sorting (FACS) should be applied for cell separation. Therefore, extracellularly represented cell surface markers were identified for both cell types to preserve the cells after separation for further in vitro cultivation. Butyrophilin (BTN1A1) was chosen as a marker for luminal lactocytes, as this protein is highly expressed in the secretory epithelium of the mammary gland during lactation and is located on the apical cell surface of lactocytes. For the myoepithelial cells, the oxytocin receptor (OXTR) was determined as a suitable marker, as it is also expressed during lactation and represented on the surface of the cells.

To test the suitability of BTN1A1 and OXTR as markers for separation of luminal lactocytes and myoepithelial cells by FACS, RT-qPCR will be performed in the next step to initially determine the presence and quantity of these proteins at transcriptional level.