Establishment and Characterization of First Trimester Human Trophoblast Cells with Extended Lifespan

Graham, Charles H.; Hawley, Teresa S.; Hawley, Robert; MacDougall, John R.; Kerbel, Robert S.; Khoo, Nelson K.S.; Lala, Peeyush K.

doi:10.1006/excr.1993.1139

Cited by 902 publications

(672 citation statements)

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“…SGHPL-5 cells, exhibiting properties of extravillous cytotrophoblasts, were cultivated in a 1:1 mixture of DMEM/Ham's F-12 supplemented with 10% FCS (GibcoBRL Life Technologies, Paisley, UK) as described [18]. Trophoblastic HTR-8/SVneo cells were cultivated in RPMI 1640 (GibcoBRL) plus 5% FCS as mentioned [19].…”

Section: Cultivation Of Primary Trophoblasts and Trophoblastic Cell Lmentioning

confidence: 99%

Complex Expression Pattern of Wnt Ligands and Frizzled Receptors in Human Placenta and its Trophoblast Subtypes

et al. 2007

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Canonical Wingless (Wnt) signalling provoked by exogenous and endogenous Wnt ligands was recently shown to play a crucial role in the invasive differentiation of human trophoblasts. To gain insights into the expression pattern of the developmental regulators, we analysed all human Wnt ligands and their frizzled (FZD) receptors in the human placenta and different trophoblast model systems using semi-quantitative PCR. Fourteen out of 19 Wnt ligands and 8 out of 10 FZD receptors were detectable in placental tissues, however, expression patterns varied with gestational age and between different trophoblast subtypes suggesting cell-specific functions. Besides Wnt ligands acting through the canonical pathway, non-canonical ligands such as Wnt-5a, which may also activate alternative Wnt signalling pathways or inhibit canonical Wnt signalling, could be identified. Western blot analyses revealed secretion of Wnt-5a from primary trophoblast cultures and trophoblastic cell lines. To evaluate the potential role of Wnt-5a, SGHPL-5 trophoblast cells were transfected with luciferase reporter plasmids harbouring eight T-cell factor (TCF) DNArecognition sequences which are exclusively activated through the canonical Wnt signalling pathway. Luciferase assays revealed that Wnt-3a-induced reporter activity was repressed by recombinant Wnt-5a indicating an antagonistic role in trophoblasts. The data suggest that a complex network of Wnt ligands and FZD receptors may regulate developmental processes of the human placenta.

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Section: Cultivation Of Primary Trophoblasts and Trophoblastic Cell Lmentioning

confidence: 99%

Complex Expression Pattern of Wnt Ligands and Frizzled Receptors in Human Placenta and its Trophoblast Subtypes

et al. 2007

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“…This is not unexpected as PAI-1 expression in normal placenta throughout gestation is localized to extravillous trophoblast [9], the cell-type from which the HTR-8/SVneo cell line is derived [30]. Conversely, PAI-2 was primary localized to the syncytial layer of villous trophoblast across gestation [9,[14][15][16].…”

Section: Discussionmentioning

confidence: 99%

“…They are a cell line obtained following immortalization of a first trimester short-lived extravillous trophoblast cell line HTR-8 after transfection with SV40 large T antigen followed by selection for neomycin resistance [30]. These cells share the phenotype and function of HTR-8 cells including responsiveness to TGF-β as well as the absence of a transformed phenotype (e.g.…”

Section: Methodsmentioning

confidence: 99%

“…These cells share the phenotype and function of HTR-8 cells including responsiveness to TGF-β as well as the absence of a transformed phenotype (e.g. anchorage-independent growth or tumorigenicity in nude mice) [30,35]. In the present study, HTR-8/SVneo cells were maintained and passaged in RPMI medium supplemented with 5% fetal calf serum and 1 mM glutamine (i.e.…”

Section: Methodsmentioning

confidence: 99%

“…It is well established that siRNAs (19−21bp RNA fragments) suppress gene expression following binding to specific complementary mRNA, thereby promoting their degradation [29]. HTR-8/SVneo cells, derived from first trimester placenta, express an extravillous trophoblast phenotype and maintain characteristics of the non-immortalized HTR-8 parent cell line including proliferative and invasive potential as well as hormone responsiveness [30]. In addition, our group and others have previously demonstrated that PAI-1 expression in this cell line was induced by hypoxic treatment as well as by glucocorticoid and TGF-β [22,31].…”

Section: Introductionmentioning

confidence: 99%

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Role of Hypoxia-Inducible Transcription Factors 1α and 2α in the Regulation of Plasminogen Activator Inhibitor-1 Expression in a Human Trophoblast Cell Line

2007

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The plasminogen activator inhibitors (PAIs) play critical roles in regulating hemostatic and invasive functions of trophoblasts through suppression of plasmin-dependent fibrinolysis and extracellular matrix degradation. The expression of PAI-1 is increased under hypoxic conditions, although the mechanism remains incompletely understood. In the current study we used HTR-8/SVneo cells, a first trimester extravillous trophoblast cell line, and siRNA technology to examine the role of hypoxia-inducible transcription factors (HIFs)−1α and −2α in the regulation of PAI-1 expression. Using serum-containing and serum-free media culture media it was initially noted that levels of PAI-1, but not PAI-2 protein, were markedly induced by hypoxic (2−3% oxygen) treatment. Under hypoxic conditions, Western blotting revealed that the presence of siRNAs to HIF-1α and HIF-2α suppressed expression of their respective proteins, whereas treatment with non-targeting and cyclophilin B siRNAs did not. Importantly, incubation with siRNA to HIF-1α or HIF-2α alone reduced PAI-1 protein levels to a similar extent, with the combined treatment inducing a more profound effect. The presence of HIF siRNAs reduced levels of PAI-1 mRNA as measured by quantitative real-time PCR, indicating that HIF-1α and HIF-2 α regulate PAI-1 expression at a transcriptional level. These results indicate that both HIF-1α and HIF-2α play important and similar roles in hypoxia-mediated stimulation of PAI-1 expression in HTR-8/SVneo cells. Our findings provide insight into the physiological regulation of trophoblast PAI-1 expression in early pregnancy when placental oxygen levels are low, as well as a mechanism for over-expression of placental PAI-1 noted in pregnancies with preeclampsia.

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SV40 Tag transformation of the normal invasive trophoblast results in a premalignant phenotype. I. Mechanisms responsible for hyperinvasivess and resistance to anti-invasive action of TGFβ

et al. 1998

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Establishment and Characterization of First Trimester Human Trophoblast Cells with Extended Lifespan

Cited by 902 publications

References 0 publications

Complex Expression Pattern of Wnt Ligands and Frizzled Receptors in Human Placenta and its Trophoblast Subtypes

Complex Expression Pattern of Wnt Ligands and Frizzled Receptors in Human Placenta and its Trophoblast Subtypes

Role of Hypoxia-Inducible Transcription Factors 1α and 2α in the Regulation of Plasminogen Activator Inhibitor-1 Expression in a Human Trophoblast Cell Line

SV40 Tag transformation of the normal invasive trophoblast results in a premalignant phenotype. I. Mechanisms responsible for hyperinvasivess and resistance to anti-invasive action of TGFβ

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