The newly established bovine endometrial gland cell line (BEGC) forms gland acini in vitro and is only IFNτ-responsive (MAPK42/44 activation) after E 2 and P 4 -pre-incubation

• Published in: Placenta (Eastbourne) 2018-07, Vol.67, p.61-69
• Main Authors: Haeger, Jan-Dirk, Loch, Christian, Pfarrer, Christiane
• Format: Article
• Language: English
• Subjects: Acinar Cells - cytology; Acinar Cells - drug effects; Acinar Cells - physiology; Adnexa Uteri - cytology; Adnexa Uteri - drug effects; Adnexa Uteri - physiology; Animals; Cattle; Cell Culture Techniques; Cell Differentiation - drug effects; Cell Line; Endometrium - cytology; Enzyme Activation - drug effects; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - physiology; Estradiol - pharmacology; Female; Interferon Type I - pharmacology; MAP Kinase Signaling System - drug effects; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinase 3 - metabolism; Pregnancy Proteins - pharmacology; Progesterone - pharmacology
• ISSN: 0143-4004; 1532-3102
• DOI: 10.1016/j.placenta.2018.05.009

Uterine glands (UG) are crucial for the establishment of ruminant pregnancy and influenced (orchestrated manner) by estrogen (E ), progesterone (P ) and interferon tau (IFNτ). In the study we established a bovine endometrial glandular cell line (BGEC) and tested its functional reactivity (signaling) to IFNτ. BGEC was characterized by light microscopy (LM), epithelial markers (ezrin, CK18) [immunofluorescence (IF)/immunohistochemistry (IHC)] and ultrastructure (TEM/SEM) (apical microvilli). In vitro formation of gland acini and transepithelial-electric-resistance (TEER) measurements (EVOM) were done. The expression of mRNA-transcripts (RT-PCR) of steroid receptors (PR, PGRMC1/2, ESR1/2) and the IFNτ-system (IFNAR1/2, IRF1, 2, 9) was checked. BEGC was stimulated with IFNτ (10 ng/ml;1000 ng/ml) (15 min) after steroid pre-treatment [10 pg/ml E (two days)/20 ng/ml P (two days)]. Activation of MAPK42/44;STAT1 was evaluated (densitometrical Western Blot). BGEC cells expressed epithelial markers and possessed apical microvilli. High TEER-values could be measured (2320-2620 ohm/cm ). The assembled BEGC acini (25 days) were similar to UG in vivo (markers/ultrastructure). All transcripts (steroid receptors/IFNτ-system) could be detected in BEGC (mRNA). MAPK42/44 were significantly activated after E /P pre-treatment and IFNτ stimulation (10 ng/ml) (p