Characterisation of an in vitro system to study maternal communication with spermatozoa

2012 ◽  
Vol 24 (7) ◽  
pp. 988 ◽  
Author(s):  
Ahmed Aldarmahi ◽  
Sarah Elliott ◽  
Jean Russell ◽  
Thomas Klonisch ◽  
Sabine Hombach-Klonisch ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
In Vitro Model ◽  
Epithelial Cell Line ◽  
Prostaglandin E ◽  
Sperm Viability ◽  
Cell Passage ◽  
In Vitro System ◽  
Vitro Model

In vivo, gamete maturation, fertilisation and early embryonic development take place inside the oviduct. Several studies have indicated that local responses towards gametes and embryos are generated by the maternal reproductive tract. However, no defined in vitro model currently exists to allow detailed and systematic investigation of maternal communications with gametes and embryos. Therefore, we characterised an in vitro model based on the interaction of boar spermatozoa with an immortalised porcine oviduct epithelial cell line to evaluate different factors that may affect this model. The factors tested were sperm viability, source of spermatozoa, cell passage effect and the effect of reproductive and non-reproductive epithelial cells in the interaction with spermatozoa. After 24 h of co-incubation, RNA was extracted and used to synthesise cDNA for quantitative real-time PCR. Alteration in the expression of genes such as adrenomedullin, heat-shock 70-kDa protein 8 and prostaglandin E synthase was considered as the end point of this assay. The results showed that sperm viability and cell passage number had an effect on oviductal gene expression in response to spermatozoa. Oviductal cells showed significant alterations in gene expression when compared with non-reproductive epithelial cells. The simple in vitro system described here has potential application for further studies in our understanding of mechanisms involved in maternal interactions with spermatozoa.

scholarly journals Hypoxia Induces Mesenchymal Gene Expression in Renal Tubular Epithelial Cells: An in vitro Model of Kidney Transplant Fibrosis

Nephron Extra ◽  
2013 ◽  
Vol 3 (1) ◽  
pp. 50-58 ◽  
Author(s):  
Stephanie Zell ◽  
Roland Schmitt ◽  
Sandra Witting ◽  
Hans H. Kreipe ◽  
Kais Hussein ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Kidney Transplant ◽  
In Vitro Model ◽  
Tubular Epithelial Cells ◽  
Renal Tubular Epithelial Cells ◽  
Vitro Model ◽  
Renal Tubular

Iron-related transcriptomic variations in CaCo-2 cells, an in vitro model of intestinal absorptive cells

2006 ◽  
Vol 26 (1) ◽  
pp. 55-67 ◽  
Author(s):  
Céline Chicault ◽  
Bertrand Toutain ◽  
Annabelle Monnier ◽  
Marc Aubry ◽  
Patricia Fergelot ◽  
...  
Keyword(s):  
Gene Expression ◽  
Functional Annotation ◽  
In Vitro Model ◽  
Iron Absorption ◽  
Intracellular Iron ◽  
Absorptive Cells ◽  
Number Of Genes ◽  
Vitro Model ◽  
First Time

Regulation of iron absorption by duodenal enterocytes is essential for the maintenance of homeostasis by preventing iron deficiency or overload. Despite the identification of a number of genes implicated in iron absorption and its regulation, it is likely that further factors remain to be identified. For that purpose, we used a global transcriptomic approach, using the CaCo-2 cell line as an in vitro model of intestinal absorptive cells. Pangenomic screening for variations in gene expression correlating with intracellular iron content allowed us to identify 171 genes. One hundred nine of these genes are clustered into five types of expression profile. This is the first time that most of these genes have been associated with iron metabolism. Functional annotation of these five clusters indicates potential links between the immune response, proteolysis processes, and iron depletion. In contrast, iron overload is associated with cellular metabolism, especially that of lipids and glutathione involving redox function and electron transfer.

scholarly journals An in vitro model of erythroid egress in bone marrow

Blood ◽  
1986 ◽  
Vol 68 (1) ◽  
pp. 250-257 ◽  
Author(s):  
RE Waugh ◽  
M Sassi
Keyword(s):  
Bone Marrow ◽  
In Vitro Model ◽  
Physical Parameters ◽  
In Vitro System ◽  
Wafer Thickness ◽  
Pore Cells ◽  
A Cell ◽  
Vitro Model ◽  
Passage Times

Abstract An in vitro system has been developed that mimics the passage of erythrocytes from the bone marrow to the circulation. Bone marrow egress and its proper regulation are vital physiologic processes. However, because of the inaccessibility of the marrow, it is difficult to evaluate the various factors important in controlling these processes or even to define the precise mechanism by which egress occurs. The in vitro system has been designed to evaluate the importance of different physical parameters in regulating egress. It consists of a thin silicon wafer (thickness approximately equal to 1.0 micron) cemented over the tip of a large (15.0 micron ID) micropipette. The wafer contains a single circular pore. Cells were observed under the microscope as they passed through the pore under controlled pressures. The rate and duration of passage were obtained from videorecordings of the experiment. The measured passage times agreed well with the predictions of a simple analytical model of a cell passing through a thin aperture. The experimental results confirm the conclusion reached from the analysis that the pressures needed to drive a cell through the pore are well within the physiologic range, and the time needed to complete egress is typically less than 1.0 seconds. These results support the hypothesis that erythrocyte egress may be driven by a hydrostatic pressure difference across the pore.

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