Ultrastructure of Choriocarcinoma in Vitro

1969 ◽  
Vol 27 ◽  
pp. 362-363
Author(s):  
John C. Garancis ◽  
R. A. Pattillo
Keyword(s):  
Cell Line ◽  
Physiological Function ◽  
Cell System ◽  
Bewo Cell ◽  
Bewo Cells ◽  
Gestational Choriocarcinoma ◽  
Bewo Cell Line

Growth of cell system (BeWo-cell line) derived from human gestational choriocarcinoma has been established and continuously maintained in-vitro. Furthermore, it is evident from the previous studies that this cell line has retained the physiological function of the placental trophoblasts, namely the synthesis of human chorionic gonadotrophil(HCG).The BeWo cells were relatively small and possessed single nuclei, thus indicating that this cell line consists exclusively of cytotrophoblasts. In some instances cells appeared widely separated and their lateral surfaces were provided with numerous microvilli (Fig.1).

scholarly journals Permeability properties of monolayers of the human trophoblast cell line BeWo

1997 ◽  
Vol 273 (5) ◽  
pp. C1596-C1604 ◽  
Author(s):  
Fei Liu ◽  
Michael J. Soares ◽  
Kenneth L. Audus
Keyword(s):  
Linoleic Acid ◽  
Cell Line ◽  
Morphological Changes ◽  
Molecular Size ◽  
Bewo Cell ◽  
Human Trophoblast ◽  
In Vitro System ◽  
Size Dependent ◽  
Bewo Cell Line

The BeWo cell line (b30 clone) has been examined as a potential in vitro system to study transplacental transport. At the light and electron microscope level, the cells were observed to form confluent monolayers on polycarbonate filters in ∼5 days and morphologically resembled the typical human trophoblast. BeWo monolayers developed a modest transepithelial electrical resistance and a molecular size-dependent permeability to hydrophilic passive diffusion markers, fluorescein, and selected fluorescein-labeled dextrans. Linoleic acid permeation across BeWo monolayers was asymmetric, saturable, and inhibited by low temperature and excess competing fatty acid. Forskolin and 8-bromoadenosine 3′,5′-cyclic monophosphate treatments stimulated morphological changes in BeWo cultures and enhanced the asymmetric passage of linoleic acid across the BeWo monolayers while having minimal effects on passive permeability, affirming that the differentiation state of the cells can influence membrane transporters and transmonolayer permeability. The basic permeability properties of the BeWo monolayers suggest that the cells grown on permeable supports may be examined as a convenient in vitro system to evaluate some transplacental transport mechanisms.

Transcriptomic profiling of trophoblast fusion using BeWo and JEG-3 cell lines

2019 ◽  
Vol 25 (12) ◽  
pp. 811-824 ◽  
Author(s):  
H Msheik ◽  
S El Hayek ◽  
M Furqan Bari ◽  
J Azar ◽  
W Abou-Kheir ◽  
...  
Keyword(s):  
Cell Line ◽  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
Bewo Cell ◽  
Rt Pcr ◽  
Trophoblast Differentiation ◽  
Bewo Cells ◽  
Bewo Cell Line

Abstract In human placenta, alteration in trophoblast differentiation has a major impact on placental maintenance and integrity. However, little is known about the mechanisms that control cytotrophoblast fusion. The BeWo cell line is used to study placental function, since it forms syncytium and secretes hormones after treatment with cAMP or forskolin. In contrast, the JEG-3 cell line fails to undergo substantial fusion. Therefore, BeWo and JEG-3 cells were used to identify a set of genes responsible for trophoblast fusion. Cells were treated with forskolin for 48 h to induce fusion. RNA was extracted, hybridised to Affymetrix HuGene ST1.0 arrays and analysed using system biology. Trophoblast differentiation was evaluated by real-time PCR and immunocytochemistry analysis. Moreover, some of the identified genes were validated by real-time PCR and their functional capacity was demonstrated by western blot using phospho-specific antibodies and CRISPR/cas9 knockdown experiments. Our results identified a list of 32 altered genes in fused BeWo cells compared to JEG-3 cells after forskolin treatment. Among these genes, four were validated by RT-PCR, including salt-inducible kinase 1 (SIK1) gene which is specifically upregulated in BeWo cells upon fusion and activated after 2 min with forskolin. Moreover, silencing of SIK1 completely abolished the fusion. Finally, SIK1 was shown to be at the center of many biological and functional processes, suggesting that it might play a role in trophoblast differentiation. In conclusion, this study identified new target genes implicated in trophoblast fusion. More studies are required to investigate the role of these genes in some placental pathology.

Expression of interleukin-6 (IL-6), signal transducer and activator of transcription-3 (STAT-3) and telomerase in choriocarcinomas

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Luciana Pietro ◽  
Fátima Bottcher-Luiz ◽  
Lício Augusto Velloso ◽  
Joseane Morari ◽  
Marcelo Nomura ◽  
...  
Keyword(s):  
Cell Transformation ◽  
First Trimester ◽  
Bewo Cell ◽  
Placental Development ◽  
Trimester Abortion ◽  
Bewo Cells ◽  
Culture Cells ◽  
Bewo Cell Line ◽  
Blastocyst Implantation ◽  
First Trimester Of Pregnancy

Abstract Blastocyst implantation and neoplastic invasion have some common properties related to tissue invasion, mediated by various cytokines. Aim To compare the expression of IL-6, STAT-3 and telomerase in material of abortions in the first trimester of pregnancy, at term placentas and in choriocarcinomas. Methods Immunohistochemical reactions were performed on formalin fixed and included in paraffin samples from 3 groups: abortions, normal at term placentas and choriocarcinomas. Western Blot and Real-Time PCR assays were performed on fresh material from BeWo cell line and in primary culture cells of normal placenta. Results Immunohistochemical reactions: IL-6 expression was moderate in the first trimester abortion samples and high in at term placentas and choriocarcinomas. STAT-3 was strongly positive in all groups. Telomerase expression was absent in normal at term placentas but was increased in BeWo cells. Conclusion IL-6 and STAT-3 are present in the invasion process of the normal placental development and they are maintained during the malignant transformation to choriocarcinoma. The intense telomerase expression observed in BeWo cells was strongly associated with the malignant phenotype, confirming it as a good marker for cell transformation and tumor progression.

442. Functional perspectives of caspase-14 in the human placenta

2008 ◽  
Vol 20 (9) ◽  
pp. 122
Author(s):  
L. J. White ◽  
A. K. Charles ◽  
A. M. Dharmarajan
Keyword(s):  
Cell Line ◽  
Epidermal Differentiation ◽  
Bewo Cell ◽  
Trophoblast Differentiation ◽  
Cytokeratin 18 ◽  
Bewo Cell Line ◽  
Barrier Formation ◽  
Β Hcg ◽  
Protein Transcription ◽  
Insight Into

The functional barrier for exchange between the mother and fetus in the placenta is created by the fusion of cytotrophoblasts with one another to form a continuous, multinuclear syncytiotrophoblast, which is maintained by the incorporation of underlying proliferative cytotrophoblasts. Disruption to this process has been hypothesised to be involved in the aetiology of preeclampsia. Recently we investigated caspase-14 in the context of trophoblast differentiation as it is crucial to epidermal differentiation and keratin stabilisation, revealing disparate expression of caspase-14 in the differentiating BeWo cell line. Consequently, further examination as to its functional role in trophoblast differentiation was conducted using RNA Interference (RNAi), with the hypothesis that differentiation would be suppressed following caspase-14 silencing. 100nM siRNA were delivered into the BeWo cell line for 16 h before the addition of 20µM Forskolin. Cultures were incubated for a further 24, 48 or 72 h before the extraction of RNA and protein. Transcription of the trophoblast hormone β-hCG, the endothelial mediator of eNOS, and cytokeratin 18 were found to be increased after both 24 and 48 h of differentiation following silencing, implicating caspase-14 in the regulation of these pathways. As both β-hCG and eNOS are significantly increased with trophoblast differentiation, this indicates that caspase-14 functionally suppresses BeWo differentiation. Furthermore, the differential expression of cytokeratin 18 indicates a conserved role for caspase-14 in keratin homeostasis in barrier formation. In conclusion, the suppression of caspase-14 in the BeWo cell line resulted in augmented differentiation, a trait often observed in preeclampsia. Further investigation of caspase-14 activity would provide important insight into mechanisms of trophoblast differentiation, particularly in relation to preeclampsia.

The BeWo cell line derived from a human placental choriocarcinoma is permissive for respiratory syncytial virus infection

Virus Genes ◽  
2019 ◽  
Vol 55 (3) ◽  
pp. 406-410 ◽  
Author(s):  
M. A. Velázquez-Cervantes ◽  
M. Martínez-Castillo ◽  
L. D. González-García ◽  
T. A. Vargas-Pavía ◽  
M. G. Martínez-Salazar ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Virus Infection ◽  
Cell Line ◽  
Respiratory Syncytial Virus Infection ◽  
Bewo Cell ◽  
Bewo Cell Line ◽  
Syncytial Virus

scholarly journals Cytotoxicity of Endocytosis and Efflux Inhibitors in the BeWo Cell Line

2017 ◽  
Vol 17 (5) ◽  
pp. 1-7 ◽  
Author(s):  
Mansi Shah ◽  
Luke Bourner ◽  
Shariq Ali ◽  
Sanaalarab Al-Enazy ◽  
Erik Rytting
Keyword(s):  
Cell Line ◽  
Bewo Cell ◽  
Bewo Cell Line

scholarly journals The choriocarcinoma cell line BeWo: syncytial fusion and expression of syncytium-specific proteins

Reproduction ◽  
2010 ◽  
Vol 140 (5) ◽  
pp. 759-766 ◽  
Author(s):  
Kristina Orendi ◽  
Martin Gauster ◽  
Gerit Moser ◽  
Hamutal Meiri ◽  
Berthold Huppertz
Keyword(s):  
Protein Expression ◽  
Cell Line ◽  
Cell Fusion ◽  
Cell Counting ◽  
Bewo Cell ◽  
Placental Alkaline Phosphatase ◽  
Bewo Cells ◽  
Villous Trophoblast ◽  
Choriocarcinoma Cell Line

Fusion of the trophoblast-derived choriocarcinoma cell line BeWo can be triggered by forskolin. BeWo cells are regularly used as a cell culture model to mimic in vivo syncytialisation of placental villous trophoblast. The β subunit of human chorionic gonadotropin (CGB), placental alkaline phosphatase as well as placental protein 13 (PP13, LGALS13) are exclusively expressed in the syncytiotrophoblast of the human placenta, and CGB is commonly used as a marker of syncytial differentiation. Here we tested the hypothesis that syncytial fusion precedes CGB and LGALS13 expression in trophoblast-derived BeWo cells. BeWo cells were cultured for 48 h in the presence or absence of forskolin and varying concentrations of H-89, a protein kinase A inhibitor that interferes with the forskolin-mediated pathway of syncytial fusion. LGALS13 and CGB expression were quantified by DELFIA and real-time PCR. Cell fusion was determined by morphological analysis and cell counting after immunofluorescence staining. In forskolin-stimulated BeWo cells that were hindered to fuse by treatment with H-89, levels of CGB protein expression were not altered, while LGALS13 protein and mRNA expression decreased significantly to control levels without forskolin. The LGALS13 protein expression data coincided with a significant decrease in syncytial fusion, while CGB protein expression was unaffected by rates of cell fusion and proliferation. We postulate that CGB protein expression is not necessarily linked to syncytial fusion, and thus CGB should be used with great caution as a marker of BeWo cell fusion.

scholarly journals Apical and basolateral transferrin receptors in polarized BeWo cells recycle through separate endosomes.

1991 ◽  
Vol 114 (6) ◽  
pp. 1149-1158 ◽  
Author(s):  
D P Cerneus ◽  
A van der Ende
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Cell Line ◽  
Apical Cell ◽  
Bewo Cell ◽  
Transferrin Receptors ◽  
Membrane Domains ◽  
Bewo Cells ◽  
Early Endosomes ◽  
Plasma Membrane Domains

Contrary to most other epithelia, trophoblasts in the human placenta, which form the physical barrier between the fetal and the maternal blood circulation, express high numbers of transferrin receptors on their apical cell surface. This study describes the establishment of a polarized trophoblast-like cell line BeWo, which exhibit a high expression of transferrin receptors on the apex of the cells. Cultured on permeable filter supports, BeWo cells formed a polarized monolayer with microvilli on their apical cell surface. Across the monolayer a transepithelial resistance developed of approximately 600 omega.cm2 within 4 d. Depletion of Ca2+ from the medium decreased the resistance to background levels, showing its dependence on the integrity of tight junctions. Within the same period of time the secretion of proteins became polarized. In addition, the compositions of integral membrane proteins at the apical and basolateral plasma membrane domains were distinct as determined by domain-selective iodination. Similar to placental trophoblasts, binding of 125I-labeled transferrin to BeWo monolayers revealed that the transferrin receptor was expressed at both plasma membrane domains. Apical and basolateral transferrin receptors were found in a 1:2 surface ratio and exhibited identical dissociation constants and molecular weights. After uptake, transferrin recycled predominantly to the domain of administration, indicating separate recycling pathways from the apical and basolateral domain. This was confirmed by using diaminobenzidine cytochemistry, a technique by which colocalization of endocytosed 125I-labeled and HRP-conjugated transferrin can be monitored. No mixing of the two types of ligands was observed, when both ligands were simultaneously internalized for 10 or 60 min from opposite domains, demonstrating that BeWo cells possess separate populations of apical and basolateral early endosomes. In conclusion, the trophoblast-like BeWo cell line can serve as a unique model to compare the apical and basolateral endocytic pathways of a single ligand, transferrin, in polarized epithelial cells.

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