Genes involved in angiogenesis and circulatory system development are differentially expressed in porcine epithelial oviductal cells during long-term primary in vitro culture – a transcriptomic study

Abstract

An oviduct is an essential organ for gamete transport, oocyte maturation, fertilization, spermatozoon capacitation and early embryo development. The epithelium plays an important role in oviduct functioning. The products of secretory cells provide an optimal environment and influence gamete activities and embryonic development. The oviduct physiology changes during the female cycle, thus, the ratio of the secreted molecules in the oviduct fluid differs between phases. In this study, a differential gene expression in porcine oviduct epithelial cells was examined during the long-term primary in vitro culture. The microarray expression analysis revealed 2552 genes, 1537 of which were upregulated and 995 were downregulated after 7 days of culture, with subsequent changes in expression during 30 day-long culture. The obtained genes were classified into 8 GO BP terms, connected with angiogenesis and circulatory system development, extracted by DAVID software. Among all genes, 10 most up-regulated and 10 most down-regulated genes were selected for further investigation. Interactions between genes were indicated by STRING software and REACTOME FIViz application to the Cytoscape 3.6.0 software. Most of the genes belonged to more than one ontology group. Although studied genes are mostly responsible for angiogenesis and circulatory system development, they can also be found to be expressed in processes connected with fertilization and early embryo development. The latter function is focused on more, considering the fact that these genes were expressed in epithelial cells of the fallopian tube which is largely responsible for reproductive processes.Running title: Upregulation of angiogenetic process in OEC primary cultures

Muscle Cell Morphogenesis, Structure, Development and Differentiation Processes Are Significantly Regulated during Human Ovarian Granulosa Cells In Vitro Cultivation

“…Next, the pellet with GCs was washed twice in culture medium and was centrifugated again with the same settings. The culture medium consisted of DMEM (Sigma; Merck KGaA, Darmstadt, Germany), 2% foetal bovine serum FBS (FBS; Sigma; Merck KGaA, Darmstadt, Germany), 10 mg/mL gentamicin (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 4 mm L-glutamine (stock 200 mm, Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 10,000 μg/mL streptomycin and 10,000 U/mL penicillin (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) [ 15 , 16 , 17 ].…”

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).…”

Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation

“…Each GO term and KEGG pathway was considered significantly enriched if it contained at least 5 differentially expressed genes, with p < 0.05. An RT-qPCR statistical analysis was performed using the Real Statistics Resource Pack for MS Excel 2016, also using the abovementioned methods (Microsoft Corporation, Redmond, WA, USA) [15,27,28].…”