Epithelium morphogenesis and oviduct development are regulated by significant increase of expression of genes after long-term in vitro primary culture – a microarray assays

Abstract

The correct oviductal development and morphogenesis of its epithelium are crucial factors influencing female fertility. Oviduct is involved in maintaining an optimal environment for gametes and preimplantation embryo development; secretory oviductal epithelial cells (OECs) synthesize components of oviductal fluid. Oviductal epithelium also participates in sperm binding and its hyperactivation. For better understanding of the genetic bases that underlay porcine oviductal development, OECs were isolated from porcine oviducts and established long-term primary culture. A microarray approach was utilized to determine the differentially expressed genes during specific time periods. Cells were harvested on day 7, 15 and 30 of in vitro primary culture and their RNA was isolated. Gene expression was analyzed and statistical analysis was performed. 48 differentially expressed genes belonging to “tube morphogenesis”, “tube development”, “morphogenesis of an epithelium”, “morphogenesis of branching structure” and “morphogenesis of branching epithelium” GO BP terms were selected, of which 10 most upregulated include BMP4, ARG1, SLIT2, FGFR1, DAB2, TNC, EPAS1, HHEX, ITGB3 and LOX. The results help to shed light on the porcine oviductal development and its epithelial morphogenesis, and show that after long-term culture the OECs still proliferate and maintain their tube forming properties.

Biochemical properties of cofactor and coenzyme metabolism in porcine oviductal epithelial cells – a microarray study

“…Considering the biological roles of the most up-and down-regulated genes, it can be concluded that these changes may indicate the increased metabolic and proliferation activity of studied cells in primary in vitro culture. This assumption is consistent with our earlier studies showing that, after long-term culture, the OECs still proliferate and maintain their tube forming properties [10,34].…”

“…Affymetrix microarray assays have been previously employed to investigate the expression profiles of genes involved in several important processes in oviductal epithelial cells, such as biological adhesion [7], amino acids metabolism and degradation [8], cell cycle process, proliferation [9], epithelium morphogenesis and oviduct development [10]. The aim of the present study is to further characterize the porcine oviductal epithelial cells model during long-term primary in vitro culture.…”

Genes Involved in the Processes of Cell Proliferation, Migration, Adhesion, and Tissue Development as New Potential Markers of Porcine Granulosa Cellular ProcessesIn Vitro: A Microarray Approach

“…The Affymetrix procedure was performed as previously described (Kranc et al, 2017(Kranc et al, , 2018a(Kranc et al, , 2018bBryja et al, 2018;Chamier-Gliszczyńska et al, 2018;Stefańska et al, 2018;Nawrocki et al, 2018). cDNA was reverse transcribed from total sample RNA (100 ng) (Ambion Ò WT Expression Kit).…”

Cortical Granule Distribution and Expression Pattern of Genes Regulating Cellular Component Size, Morphogenesis, and Potential to Differentiation are Related to Oocyte Developmental Competence and Maturational Capacity In Vivo and In Vitro

“…The expression of this gene was recorded in granulosa cells and theca cells in bovine, during comparative studies of these two groups of ovarian follicle cells [42,55]. The work of Stefańska et al [56] observed a high increase in DAB2 expression in oviduct epithelial cells (OECs) in primary culture, which indicated high participation of this gene in morphogenesis. Additionally, this gene is considered important for the functioning of the female reproductive system [57] and plays a role in the TGF-beta pathway as a link between TGF-beta receptors and proteins from the SMAD family [58].…”