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Transcriptomic analysis of human extravillous trophoblast cells (HTR8/Svneo) cultured as 3D spheroids and 2D monolayers

ABSTRACT: Transcriptomic analysis of human extravillous trophoblast cells (HTR8/Svneo) cultured as 3D spheroids and 2D monolayers

PROVIDER: PRJNA523498 | ENA |

REPOSITORIES: ENA

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Transcriptomic analysis of human extravillous trophoblast cells (HTR8/Svneo) cultured as 3D spheroids and 2D monolayers

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Project description:Here, we have undertaken comparative label-free quantitative (LFQ) proteomic profiling of the proteins expressed in BeWo and HTR8/SVneo cell lines using a mass spectrometry approach. We have identified 1557 proteins in total. 915 proteins were expressed in both cell lines including various transcription factors, chaperones and transport proteins. A further 338 and 304 proteins were uniquely expressed in BeWo and HTR8/SVneo cells respectively. Our bioinformatics analysis reflects the known epithelial and mesenchymal phenotypes of these cell models with principal differences in GO observed in ‘Cell junction’, ‘Catenin complex’ as well as ‘Cell adhesion’ and ‘Cell differentiation’. Our novel comparative proteomic analysis of these trophoblastic cell lines will set the stage for the use of trophoblastic cell lines for the study of pregnancy disorders and further research focusing on placental function.

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Gene expression profiling of primary human bone marrow mesenchymal stromal cells (MSCs) cultured as 2D monolayers and 3D cellular aggregates (spheroids)

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Effect of depletion of TET3 on gene expression of HTR8-SVneo trophoblast cells.

Project description:To investigate the function TET3 in the regulation of DNA methylation, we established HTR8-SVneo cell lines in which target gene has been knocked down by siRNA. We then performed gene expression profiling analysis using data obtained from RNA-seq for three replicates.

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Genomic characterization of ovarian cancer spheroids

Project description:Spheroids are 3D multi-cell aggregates formed in non-addherent culture conditions. In ovarian cancer (OC), they serve as a vehicle for cancer cell dissemination in the peritoneal cavity. We investigated genes and networks upregulated in three dimensional (3D) versus two-dimensional (2D) culture conditions by Affymetrix gene expression profiling and identified ALDH1A1, a cancer stem cell marker as being upregulated in OC spheroids. Network analysis confirmed ALDH1A1 upregulation in spheroids in direct connection with elements of the beta-catenin pathway. A parallel increase in the expression levels of beta-catenin and ALDH1A1 was demonstrated in spheroids vs. monolayers an in successive spheroid generations by using OC cell liness and primary OC cells. The percentage of Aldefluor positive cells was significantly higher in spheroids vs. monolayers in IGROV1, A2780, SKOV3, and primary OC cells. B-catenin knock-down decreased ALDH1A1 expression and chromatin immunoprecipitation demonstrated that beta-catenin directly binds to the ALDH1A1 promoter. Both siRNA mediated beta-catenin knock-down and a novel ALDH1A1 small molecule enzymatic inhibitor described here for the first time, decreased the number of OC spheroids (p<0.001) and cell viability. These data strongly support the role of beta-catenin regulated ALDH1A1 in the maintenance of OC spheroids and of a stem cell phenotype and propose new ALDH1A1 inhibitors targeting this cell population. Different gene profiles were observed in ovarian cancer spheroids versus ovarian cancer monolayers. Nine samples were analyzed in triplicate. Each group is a reference.

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Project description:Transcriptional change of HTR8/Svneo cells over expressed with PLAC8-EGFP

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SIRT1 binding proteins in human first trimester villi and HTR8/SVneo cells

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