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Gene expression profiles of trophoblast cell lines

ABSTRACT: Invasive extravillous trophoblasts (EVTs) of the human placenta are critically involved in successful pregnancy outcome since they remodel the uterine spiral arteries to increase blood flow and oxygen delivery to the placenta and the developing fetus. To gain more insights into their biological role different primary cell culture models are commonly utilised. However, access to early placental tissue may be limited and primary trophoblasts rapidly cease proliferation in vitro impairing genetic manipulation. Hence, trophoblastic cell lines have been widely used as surrogates to study EVT function. Although the cell lines share some molecular marker expression with their primary counterpart, it is unknown to what extent they recapture the invasive phenotype of EVT. Therefore, we here report the first thorough GeneChip analyses of SGHPL-5, HTR-8/SVneo, BeWo, JEG-3 and the novel ACH-3P trophoblast cells in comparison to previously analysed primary villous cytrophoblasts and extravillous trophoblasts.

ORGANISM(S): Homo sapiens

PROVIDER: GSE20510 | GEO | 2010/09/23

SECONDARY ACCESSION(S): PRJNA125227

REPOSITORIES: GEO

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Gene expression profiles of trophoblast cell lines

Project description:Invasive extravillous trophoblasts (EVTs) of the human placenta are critically involved in successful pregnancy outcome since they remodel the uterine spiral arteries to increase blood flow and oxygen delivery to the placenta and the developing fetus. To gain more insights into their biological role different primary cell culture models are commonly utilised. However, access to early placental tissue may be limited and primary trophoblasts rapidly cease proliferation in vitro impairing genetic manipulation. Hence, trophoblastic cell lines have been widely used as surrogates to study EVT function. Although the cell lines share some molecular marker expression with their primary counterpart, it is unknown to what extent they recapture the invasive phenotype of EVT. Therefore, we here report the first thorough GeneChip analyses of SGHPL-5, HTR-8/SVneo, BeWo, JEG-3 and the novel ACH-3P trophoblast cells in comparison to previously analysed primary villous cytrophoblasts and extravillous trophoblasts. To identify EVT-specific gene expression signatures in trophoblast cell lines, we calcuted differentially expressed genes between pre-defined groups based on the distinct origins of the five trophoblast cell lines under investigation. Comparison 1 comprised EVT, HTR-8/Svneo and SGHPL-5 vs choriocarcinoma cells (ACH-3P, BeWo, JEG-3). Comparison 2 comprised EVT, ACH-3P, BeWo, JEG-3 vs extravillous trophoblast cell lines (HTR-8/SVneo, SGHPL-5).

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Project description:Zika virus (ZIKV) infection at the maternal-placental interface is associated with adverse pregnancy outcomes including fetal demise and pregnancy loss. To determine how infection impacts placental trophoblasts, we utilized rhesus macaque trophoblast stem cells (TSC) that can be differentiated into early gestation syncytiotrophoblasts (ST) and extravillous trophoblasts (EVT). TSCs and STs, but not EVTs, were highly permissive to productive infection with ZIKV strain DAK AR 41524. The impact of ZIKV on the cellular transcriptome showed that infection of TSCs and STs increased expression of immune related genes, including those involved in type I and type III interferon responses. ZIKV exposure altered extracellular vesicle (EV) protein, mRNA, and miRNA cargo, regardless of productive infection. These findings suggest that early gestation macaque TSCs and STs are permissive to ZIKV infection, and that EV analysis may provide a foundation for identifying non-invasive biomarkers of placental infection in a highly translational model.

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A human maternal-fetus interface transcriptome atlas

Project description:Trophoblasts undergos dynamic differentiation and self-renewal during pregnancy. Expecially, EVTs anchor the placenta to the uterus to ensure pregnancy success, and its invasive excessiveness is a probable consequence of ruptured tubal ectopic pregnancy (REP), that resulting in maternal infertile and/or even deaths. We present REP, abortive tubal ectopic pregnancy (AEP)and intrauterine pregnancy (IP) atlas by integrating single-cell RNA-seq. We define cell subtypes within EVTs and each of the feature genes. Further, we utilize monocle3 and RNA velocty to identify the developmental trajectories of EVT subsets. CellPhoneDB and cell-chat unravel secretion of CSF1 by falloppian tubal secretory epithelial cells directly interact with CSF1R of EVT to strengthens the invasive ability.

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Project description:Transcriptional profiling comparison of human primary villous trophoblast and extra-villous trophoblast (VT and EVT respectively) cells, isolated from the placenta at 8 to 12 weeks gestation, with complimentary transcriptional profiling of choriocarcinoma cell lines JEG-3 and JAR. Based on phenotypic markers, JEG is frequently used as a model for EVT and JAR is used as a model for VT.

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An improved protocol for targeted differentiation of human iPSCs into HLA-G expressing trophoblasts cells enabling modelling of placenta-related disorders.

Project description:Abnormalities at any stage of trophoblast development may result in pregnancy-related com-plications. Many of these adverse outcomes are discovered later in pregnancy, but the underly-ing pathomechanisms constitute during the first trimester. Acquiring developmentally relevant material to elucidate the disease mechanisms is difficult. Human pluripotent stem cell (hPSC) technology can provide a renewable source of relevant cells. BMP4, A83-01, and PD173074 (BAP) treatment drives trophoblast commitment of hPSCs towards syncytiotrophoblast (STB) but lacks extravillous trophoblast (EVT) cells. EVTs mediate key functions during placentation, remodel-ing of uterine spiral arteries, and maintenance of immunological tolerance. We optimized the protocol for more efficient generation of HLA-Gpos EVT-like trophoblasts from primed hiPSCs. Increasing the concentrations of A83-01 and PD173074, while decreasing bulk cell density re-sulted in the increase of HLA-G of up to 71%. Gene expression profiling support the advance-ments of our treatment regarding the generation of trophoblast cells. The reported differentia-tion protocol will allow the on-demand access to human trophoblast cells enriched for HLA-Gpos EVT-like cells, allowing the elucidation of placenta-related disorders and to investigate the im-munological tolerance towards the fetus, overcoming the difficulties in obtaining primary EVTs without the need for a complex differentiation pathway via naïve pluripotent or trophoblast stem cells.

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Zika virus impacts extracellular vesicle composition and cellular gene expression in macaque early gestation trophoblasts

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Project description:Zika virus (ZIKV) infection at the maternal-placental interface is associated with adverse pregnancy outcomes including fetal demise and pregnancy loss. To determine how infection impacts placental trophoblasts, we utilized rhesus macaque trophoblast stem cells (TSC) that can be directed to differentiate into early gestation syncytiotrophoblasts (ST) and extravillous trophoblasts (EVT). TSCs and STs, but not EVTs, were highly permissive to productive infection with ZIKV strain DAK AR 41524. The impact of ZIKV on the cellular transcriptome showed that infection of TSCs and STs increased expression of immune related genes, including those involved in type I and type III interferon responses. ZIKV exposure altered extracellular vesicle (EV) protein, mRNA, and miRNA cargo, regardless of productive infection. These findings suggest that early gestation macaque TSCs and STs are permissive to ZIKV infection, and that EV analysis may provide a foundation for identifying non-invasive biomarkers of placental infection in a highly translational model.

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