A long‐term in vitro culture of bovine epithelial cells on collagen rafts

2021 ◽  
Author(s):  
Milagros Peña‐Zanoni ◽  
Daniela Celeste García ◽  
Mariela Roldán‐Olarte ◽  
Pablo Alberto Valdecantos
Keyword(s):  
Epithelial Cells ◽  
In Vitro Culture

scholarly journals Thymus Extracellular Matrix‐Derived Scaffolds Support Graft‐Resident Thymopoiesis and Long‐Term In Vitro Culture of Adult Thymic Epithelial Cells

2021 ◽  
pp. 2010747
Author(s):  
M. Adelaide Asnaghi ◽  
Thomas Barthlott ◽  
Fabiana Gullotta ◽  
Valentina Strusi ◽  
Anna Amovilli ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
In Vitro Culture ◽  
Thymic Epithelial Cells

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

2018 ◽  
Vol 6 (2) ◽  
pp. 39-47 ◽  
Author(s):  
Joanna Budna ◽  
Piotr Celichowski ◽  
Sandra Knap ◽  
Maurycy Jankowski ◽  
Magdalena Magas ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Epithelial Cells ◽  
In Vitro Culture ◽  
Cellular Proliferation ◽  
Inflammatory Responses ◽  
Proliferation Capacity ◽  
Genes Expression ◽  
Reproductive Events

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.

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 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 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

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

scholarly journals Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

2021 ◽  
Vol 22 (13) ◽  
pp. 6663
Author(s):  
Maurycy Jankowski ◽  
Mariusz Kaczmarek ◽  
Grzegorz Wąsiatycz ◽  
Claudia Dompe ◽  
Paul Mozdziak ◽  
...  
Keyword(s):  
Stem Cells ◽  
In Vitro Culture ◽  
Osteogenic Differentiation ◽  
Molecular Mechanisms ◽  
Marker Genes ◽  
P Value ◽  
Illumina Platform

Next-generation sequencing (RNAseq) analysis of gene expression changes during the long-term in vitro culture and osteogenic differentiation of ASCs remains to be important, as the analysis provides important clues toward employing stem cells as a therapeutic intervention. In this study, the cells were isolated from adipose tissue obtained during routine surgical procedures and subjected to 14-day in vitro culture and differentiation. The mRNA transcript levels were evaluated using the Illumina platform, resulting in the detection of 19,856 gene transcripts. The most differentially expressed genes (fold change >|2|, adjusted p value < 0.05), between day 1, day 14 and differentiated cell cultures were extracted and subjected to bioinformatical analysis based on the R programming language. The results of this study provide molecular insight into the processes that occur during long-term in vitro culture and osteogenic differentiation of ASCs, allowing the re-evaluation of the roles of some genes in MSC progression towards a range of lineages. The results improve the knowledge of the molecular mechanisms associated with long-term in vitro culture and differentiation of ASCs, as well as providing a point of reference for potential in vivo and clinical studies regarding these cells’ application in regenerative medicine.

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