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

2019 ◽  
Vol 7 (2) ◽  
pp. 77-85 ◽  
Author(s):  
Katarzyna Stefańska ◽  
Ievgenia Kocherova ◽  
Sandra Knap ◽  
Magdalena Kulus ◽  
Piotr Celichowski ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Differentially Expressed Genes ◽  
Cellular Protein ◽  
Early Embryo ◽  
Female Reproductive Tract ◽  
Differentially Expressed ◽  
In Vitro Cultivation ◽  
Protein Location

AbstractThe oviduct is a part of female reproductive tract that is essential for successful fertilization and early embryo development. It is lined with epithelium consisting of two types of cells: ciliated and secretory. The primary function of ciliated oviductal epithelial cells (OECs) is to support the transport of gametes and embryos through the ovary, whereas secretory OECs produce components of the oviductal fluid. Undoubtedly, the oviductal epithelium plays a major part in the early aspects of pregnancy development, from providing an optimal environment for gametes and embryos to supporting fertilization. Therefore, our aim was to gain a better insight into the genetic changes underlying function of these cells. We have harvested OECs from crossbred gilts (n=45), at the age of about nine months and which displayed two regular estrous cycles, and established long-term primary culture of porcine OECs. Microarray analysis was utilized to determine differentially expressed genes during day 1, 7, 15 and 30 of cultivation, with our results revealing54 differentially expressed genes belonging to three ontology groups: „maintenance of location”, „maintenance of protein location” and „maintenance of protein location in cell”. Since the biochemistry and morphology of epithelial cells may change during long term cultivation, we conclude that our results are a reflection of these changes and help to shed a light on porcine OECs properties in in vitro environment.Running title: Maintenance of cellular protein location in porcine epithelial oviductal cells

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

2018 ◽  
Vol 6 (4) ◽  
pp. 195-204 ◽  
Author(s):  
Katarzyna Stefańska ◽  
Agata Chamier-Gliszczyńska ◽  
Maurycy Jankowski ◽  
Piotr Celichowski ◽  
Magdalena Kulus ◽  
...  
Keyword(s):  
Primary Culture ◽  
Differentially Expressed Genes ◽  
Differentially Expressed ◽  
Preimplantation Embryo Development ◽  
Sperm Binding ◽  
Expression Of Genes ◽  
Tube Morphogenesis ◽  
Oviductal Fluid

AbstractThe 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.

scholarly journals “Biological Adhesion” is a Significantly Regulated Molecular Process during Long-Term Primary In Vitro Culture of Oviductal Epithelial Cells (Oecs): A Transcriptomic and Proteomic Study

2019 ◽  
Vol 20 (14) ◽  
pp. 3387 ◽  
Author(s):  
Joanna Budna-Tukan ◽  
Agata Światły-Błaszkiewicz ◽  
Piotr Celichowski ◽  
Sandra Kałużna ◽  
Aneta Konwerska ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
In Vitro Culture ◽  
Early Embryo ◽  
Maldi Tof ◽  
Biological Adhesion ◽  
Proteomic Study ◽  
Reproductive Events

Oviductal epithelial cells (OECs) actively produce stimulating and protecting factors, favoring survival and viability of gametes and early embryos. The oviduct participates in the initial reproductive events, which strongly depends on adhesion. The analysis of differential gene expression in OECs, during long-term in vitro culture, enables recognition of new molecular markers regulating several processes, including “biological adhesion”. Porcine oviducts were stained with hematoxylin and eosin, as well as with antibodies against epithelial markers. Then, OECs were long-term in vitro cultured and after 24 h, 7, 15, and 30 days of culture were subjected to transcriptomic and proteomic assays. Microarrays were employed to evaluate gene expression, with Matrix-assisted laser desorption/ionization-time of light (MALDI-TOF) mass spectrometry applied to determine the proteome. The results revealed proper morphology of the oviducts and typical epithelial structure of OECs during the culture. From the set of differentially expressed genes (DEGs), we have selected the 130 that encoded proteins detected by MALDI-TOF MS analysis. From this gene pool, 18 significantly enriched gene ontology biological processes (GO BP) terms were extracted. Among them we focused on genes belonging to “biological adhesion” GO BP. It is suggested that increased expression of studied genes can be attributed to the process of intensive secretion of substances that exhibit favorable influence on oviductal environment, which prime gametes adhesion and viability, fertilization, and early embryo journey.

scholarly journals Klebsiella pneumoniae infection causes mitochondrial damage and dysfunction in bovine mammary epithelial cells

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Jia Cheng ◽  
Jv Zhang ◽  
Jingyue Yang ◽  
Bing Yi ◽  
Gang Liu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Differentially Expressed Genes ◽  
Mammary Epithelial Cells ◽  
Mitochondrial Damage ◽  
Mammary Epithelial ◽  
Differentially Expressed ◽  
Cellular Damage ◽  
Bacterial Strains ◽  
Bovine Mammary Epithelial Cells

AbstractKlebsiella pneumoniae, an important cause of bovine mastitis worldwide, is strongly pathogenic to bovine mammary epithelial cells (bMECs). Our objective was to determine the role of mitochondrial damage in the pathogenicity of K. pneumoniae on bMECs, by assessing several classical indicators of mitochondrial dysfunction, as well as differentially expressed genes (DEGs). Two K. pneumoniae strains (HLJ-D2 and HB-AF5), isolated from cows with clinical mastitis (CM), were used to infect bMECs (MAC-T line) cultured in vitro. In whole-transcriptome analysis of bMECs at 6 h post-infection (hpi), there were 3453 up-regulated and 3470 down-regulated genes for HLJ-D2, whereas for HB-AF5, there were 2891 up-regulated and 3278 down-regulated genes (P < 0.05). Based on GO term enrichment of differentially expressed genes (DEGs), relative to the controls, the primary categories altered in K. pneumoniae-infected bMECs included cellular macromolecule metabolism, metabolic process, binding, molecular function, etc. Infections increased (P < 0.05) malondialdehyde concentrations and formation of reactive oxygen species in bMECs. Additionally, both bacterial strains decreased (P < 0.05) total antioxidant capacity in bMECs at 6 and 12 hpi. Furthermore, infections decreased (P < 0.05) mitochondrial membrane potential and increased (P < 0.01) mitochondrial calcium concentrations. Finally, severe mitochondrial swelling and vacuolation, as well as mitochondrial rupture and cristae degeneration, were detected in infected bMECs. In conclusion, K. pneumoniae infections induced profound mitochondrial damage and dysfunction in bMECs; we inferred that this caused cellular damage and contributes to the pathogenesis of K. pneumoniae-induced CM in dairy cows.

scholarly journals 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

2018 ◽  
Vol 6 (4) ◽  
pp. 163-173 ◽  
Author(s):  
Agata Chamier-Gliszczyńska ◽  
Maciej Brązert ◽  
Patrycja Sujka-Kordowska ◽  
Małgorzata Popis ◽  
Katarzyna Ożegowska ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
In Vitro Culture ◽  
Embryo Development ◽  
System Development ◽  
Circulatory System ◽  
Early Embryo ◽  
Secretory Cells ◽  
Early Embryo Development

AbstractAn 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.

scholarly journals “Cell cycle process”, “cell division” and “cell proliferation” belong to ontology groups highly regulated during long–term culture of porcine oviductal epithelial cells

2019 ◽  
Vol 7 (1) ◽  
pp. 15-24 ◽  
Author(s):  
Magdalena Kulus ◽  
Małgorzata Józkowiak ◽  
Jakub Kulus ◽  
Małgorzata Popis ◽  
Blanka Borowiec ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Division ◽  
Epithelial Cells ◽  
P Value ◽  
In Vitro Cultivation ◽  
Bioinformatics Analyses ◽  
Cell Culture Techniques

AbstractMorphological and biochemical changes in the cells surrounding the oocyte seem to be extremely important in an effective fertilization process. Thanks to advanced cell culture techniques, as well as biochemical and bioinformatics analyses, we can partly imitate the phenomena occurring in the living organism. Previous studies showed a possibility of short – and long – term OEC in vitro cultivation, during which these cells have shown to have significant proliferation and expression of genes responsible for differentiation. Our research was aimed at maintaining a culture of porcine oviduct epithelial cells and analyzing their gene expression profile. The study employed cross-bred gilts at the age of about 9 months, obtained from commercial herds. With the use of Affymetrix® Porcine Gene 1.1 ST Array Strip, we have examined the expression of 12257 transcripts. Genes with fold change higher than abs (2) and with corrected p-value lower than 0.05 were considered as differentially expressed. We chose 20 genes with the most marked expression (10 up – regulated, 10 down – regulated) for further investigation in the context of literature sources. These genes belonged to three ontological groups: “cell cycle process”, “cell division” and “cell proliferation”. The results obtained from these studies may be the basis for further molecular analyses.

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

2019 ◽  
Vol 7 (3) ◽  
pp. 125-133 ◽  
Author(s):  
Ievgeniia Kocherova ◽  
Magdalena Kulus ◽  
Claudia Dompe ◽  
Paweł Antosik ◽  
Dorota Bukowska ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
In Vitro Culture ◽  
Dynamic Structure ◽  
Biochemical Properties ◽  
Early Embryo ◽  
Metabolic Process ◽  
Affymetrix Microarray ◽  
Innermost Layer

AbstractThe oviduct is a key organ responsible for ultimate oocytes maturation, transport of gametes, sperm capacitation, fertilization, as well as early embryo development. Its innermost layer, oviductal epithelium, represents a highly dynamic structure which undergoes changes in response to different physiological and pathological processes. Previously, the expression profile of genes involved in several important processes in porcine oviductal epithelial cells (OECs) during long-term primary in vitro culture. The present study further characterizes the porcine OECs model using Affymetrix microarray assay and it analyzes gene expression changes observed on the 7th, 15th and 30th day of culture. 25 genes belonging to “coenzyme metabolic process”, “cofactor biosynthetic process” and “cofactor metabolic process” GO BP terms were differentially expressed in culture. The most up-regulated genes were ALDH1L2, P2RX7, PANK1, ACSS2, SCD, AASS and PDK3. In contrast, several genes appeared to be significantly down-regulated, e.g. ACSL4 and HAAO. Considering the biological roles of the most 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.Running title: Cofactor and coenzyme metabolism in porcine oviductal epithelial cells

scholarly journals Low-dose Drosera rotundifolia induces gene expression changes in 16HBE human bronchial epithelial cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fabio Arruda-Silva ◽  
Paolo Bellavite ◽  
Marta Marzotto
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Differentially Expressed Genes ◽  
Bronchial Epithelial Cells ◽  
Differentially Expressed ◽  
Biological Functions ◽  
Bronchial Epithelial ◽  
Cell Functions ◽  
Drosera Rotundifolia

AbstractDrosera rotundifolia has been traditionally used for the treatment of respiratory diseases in phytotherapy and homeopathy. The mechanisms of action recognized so far are linked to the known effects of specific components, such as flavonoids, but are not completely understood. In this study, the biological functions of D. rotundifolia were explored in vitro following the treatment of bronchial epithelial cells, which are the potential targets of the pharmacological effects of the herbal medicine. To do so, the whole plant ethanolic extract was 1000-fold diluted in water (D. rotundifolia 3×) and added to a 16HBE human cell line culture for 3 h or 6 h. The effects on gene expression of the treatments and corresponding controls were then investigated by RNA sequencing. The differentially expressed genes were validated through RT-qPCR, and the enriched biological functions involved in the effects of treatment were investigated. D. rotundifolia 3× did not impair cell viability and was shown to be a stimulant of cell functions by regulating the expression of dozens of genes after 3 h, and the effects were amplified after 6 h of treatment. The main differentially expressed genes encoded ligands of epithelial growth factor receptor, proteins involved in xenobiotic detoxification and cytokines, suggesting that D. rotundifolia 3× could stimulate self-repair systems, which are impaired in airway diseases. Furthermore, D. rotundifolia 3× acts on a complex and multifaceted set of genes and may potentially affect different layers of the bronchial mucosa.

Human thymic epithelial cells maintain long-term survival of clonogenic myeloid and erythroid progenitor cells in vitro

2000 ◽  
Vol 111 (1) ◽  
pp. 363-370 ◽  
Author(s):  
Katsuto Takenaka ◽  
Mine Harada ◽  
Tomoaki Fujisaki ◽  
Koji Nagafuji ◽  
Shinichi Mizuno ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Progenitor Cells ◽  
Thymic Epithelial Cells ◽  
Erythroid Progenitor ◽  
Term Survival ◽  
Long Term Survival ◽  
Erythroid Progenitor Cells
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