Cultured bovine granulosa cells rapidly lose important features of their identity and functionality but partially recover under long-term culture conditions.
Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.
Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany.
Vengala Rao
VR
Yenuganti
Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany. vanselow@fbn-dummerstorf.de.
Jens
J
Vanselow
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D000074842:Forkhead Box Protein L2 / Q000235:genetics / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology* / Q000378:metabolism*; D006728:Hormones / Q000096:biosynthesis; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D012708:Sertoli Cells / Q000166:cytology / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis; D013997:Time Factors
D015415:Biomarkers; D000074842:Forkhead Box Protein L2; D006728:Hormones; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D013256:Steroids
Cell identity; Cell plating; FOXL2; PTGS2; SOX9