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Ontology groups representing angiogenesis and blood vessels development are highly up-regulated during porcine oviductal epithelial cells long-term real-time proliferation – a primary cell culture approach

, Zabel M, Kempisty B. Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro a signaling pathways activation approach. Med J Cell Biol. 2018;6:18–26; DOI:10.2478/acb-2018-0004. 10. Ferrara N. Vascular Endothelial Growth Factor: Basic Science and Clinical Progress. Endocr Rev. 2004;25:581–611; DOI:10.1210/er.2003-0027. 11. Ribeiro LA, Bacci ML, Seren E, Tamanini C, Forni M. Characterization and differential expression of vascular endothelial growth factor isoforms and receptors in swine

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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.

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Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro – a signaling pathways activation approach

Abstract

The ovary is part of the reproductive system, possessing very important functions in the reproduction process (ovum and embryo transfer, providing a suitable environment for sperm capacitation, etc.). There are two types of cells in the fallopian tubes: alveolar and secretive cells. These study shows the metabolic processes in pig oviductal epithelial cells associated with the activation of signaling pathways of amino acids metabolism and degradation during long-term in vitro culture. Oviductal epithelial cells from 45 colonies in the anestrous phase of the estrous cycle have been utilized in this study. RNA extract from the OEC primary cultures was pooled after 24h, 7days, 15 days and 30 days from the beginning of culture and the transcriptome investigated by Affymetrix® Porcine Gene 1.1 ST. From the whole transcript that consisted of 2009 different genes, 1537 were upregulated and 995 were downregulated after 7 days of culture, 1471 were upregulated and 1061 were downregulated after 15 days of culture and 1329 were upregulated and 1203 were downregulated after 30 days of culture. The results of these studies provide, for the first time, information on the activation of metabolic pathways of amino acids such as valine, leucine, isoleucine, cysteine, and methionine in the investigated tissue. They also indicate genes that may be OECs-specific genetic markers that are expressed or upregulated during long-term in vitro culture.

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Cation homeostasis and transport related gene markers are differentially expressed in porcine buccal pouch mucosal cells during long-term cells primary culture in vitro

, Kempisty B. New gene markers for metabolic processes and homeostasis in porcine buccal pouch mucosa during cells long term-cultivation - a primary culture approach. IJMS. 2018;19(4):1027; DOI:10.3390/ijms19041027. 7. Huang DW, Sherman BT, Tan Q, Collins JR, Alvord WG, Roayaei J, Stephens R, Baseler MW, Lane HC, Lempicki RA. The DAVID Gene Functional Classification Tool: a novel biological module-centric algorithm to functionally analyze large gene lists. Genome Biol. 2007;8(9):R183; DOI:10.1186/gb-2007-8-9-r183. 8. Mering C von, Jensen LJ, Snel B, Hooper SD

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Genes regulating biochemical pathways of oxygen metabolism in porcine oviductal epithelial cells during long-term IVC

epithelial cells (OECs) primary culture in vitro – a signaling pathways activation approach. Med J Cell Biol. 2018;6:18–26; DOI:10.2478/acb-2018-0004. 10.2478/acb-2018-0004 Kranc W Jankowski M Budna J Celichowski P Khozmi R Bryja A Borys S Dyszkiewicz-Konwińska M Jeseta M Magas M Bukowska D Antosik P Brüssow KP Bruska M Nowicki M Zabel M Kempisty B Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro – a signaling pathways

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The genes regulating maintenance of cellular protein location are differentially expressed in porcine epithelial oviductal cells during longterm in vitro cultivation

fertilization, our aim was to gain a better insight into genetic changes that underlie these cells’ function. We established a long-term primary culture of porcine OECs and performed microarray analysis, checking gene expression in specific time periods. Our results revealed a set of genes belonging to “maintenance of location”, “maintenance of protein location in cell” and “maintenance of protein location” that were differentially expressed, indicating that they may be possible marker genes of biochemical and morphological changes in OECs. Material and Methods Animals

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Fatty Acids Related Genes Expression Undergo Substantial Changes in Porcine Oviductal Epithelial Cells During Long-Term Primary Culture

Abstract

The process of reproduction requires several factors, leading to successful fertilization of an oocyte by a single spermatozoon. One of them is the complete maturity of an oocyte, which is acquired during long stages of folliculogenesis and oogenesis. Additionally, the oviduct, composed of oviductal epithelial cells (OECs), has a prominent influence on this event through sperm modification and supporting oocyte’s movement towards uterus. OECs were isolated from porcine oviducts. Cells were kept in primary in vitro culture for 30 days. After 24h and on days 7, 15 and 30 cells were harvested, and RNA was isolated. Transcript changes were analyzed using microarrays. Fatty acids biosynthetic process and fatty acids transport ontology groups were selected for analysis and described. Results of this study indicated that majority of genes in both ontology groups were up-regulated on day 7, 15 and 30 of primary in vitro culture. We analyzed genes involved in fatty acids biosynthetic process, including: GGT1, PTGES, INSIG1, SCD, ACSL3, FADS2, FADS1, ACSS2, ALOX5AP, ACADL, SYK, ACACA, HSD17B8, FADS3, OXSM, and transport, including: ABCC2, ACSL4, FABP3, PLA2G3, PPARA, SYK, PPARD, ACACA and P2RX7. Elevated levels of fatty acids in bovine and human oviducts are known to reduce proliferation capacity of OECs and promote inflammatory responses in their microenvironment. Most of measured genes could not be connected to reproductive events. However, the alterations in cellular proliferation, differentiation and genes expression during in vitro long-term culture were significant. Thus, we can treat them as putative markers of changes in OECs physiology.

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

Oviductal Epithelial Cells (Oecs): A Transcriptomic and Proteomic Study Int J Mol Sci 2019 20 3387 10.3390/ijms20143387 8 Kranc W, Jankowski M, Budna J, Celichowski P, Khozmi R, Bryja A, Borys S, Dyszkiewicz-Konwińska M, Jeseta M, Magas M, Bukowska D, Antosik P, Brüssow KP, Bruska M, Nowicki M, Zabel M, Kempisty B. Amino acids metabolism and degradation is regulated during porcine oviductal epithelial cells (OECs) primary culture in vitro – a signaling pathways activation approach. Med J Cell Biol. 2018;6:18–26; DOI:10.2478/acb-2018-0004. 10.2478/acb-2018

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Ovarian follicular cells - living in the shadow of stemness cellular competence

, Bruska M, Nowicki M, Kempisty B. Selected molecular and physiological aspects of mammalian ovarian granulosa cells in primary culture. Med Weter. 2016;72:723–7; DOI:10.21521/mw.5606. Kranc W Chachuła A Bryja A Ciesiółka S Budna J Wojtanowicz-Markiewicz K Sumelka E Borys S Antosik P Bukowska D Bruska M Nowicki M Kempisty B Selected molecular and physiological aspects of mammalian ovarian granulosa cells in primary culture Med Weter 2016 72 723 7 10.21521/mw.5606 3 Kottarathil VD, Antony MA, Nair IR

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Genes encoding proteins regulating fatty acid metabolism and cellular response to lipids are differentially expressed in porcine luminal epithelium during long-term culture

molecular mechanisms of these changes have not been discovered. Hence, understanding the metabolism of oviduct epithelial cells and the level of expression of individual groups of genes seems to be a way to deepen the knowledge about the broadly understood reproduction. The aim of our research was to maintain the culture of epithelial cells of the porcine fallopian tube in primary culture. We carried out this culture over a long term, lasting up to 30 days. We have made observations of biochemical and morphological changes occurring during the cell culture. We analyzed

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