Cytogenetic and DNA-fingerprint characterization of choriocarcinoma cell lines and a trophoblast/choriocarcinoma cell hybrid.
ABSTRACT We report the successful fusion of human choriocarcinoma cells with normal human trophoblast cells to a choriocarcinoma/trophoblast hybrid. The hybrid cells ACH1P were derived from fusion of primary male trophoblast cells with the HGPRT-defective choriocarcinoma cell line AC1-1. The karyotypes of the parental choriocarcinoma cell line JEG-3, its HGPRT-defective mutant clones AC1-1, AC1-5, and AC1-9, and the choriocarcinoma/trophoblast hybrid ACH1P are presented, together with a detailed characterization of the AC1-specific chromosomal marker add(X)(q26) using conventional cytogenetic banding techniques and multiplex-fluorescence in situ hybridization (M-FISH). To our knowledge, this is the first report of a stably proliferating human cell hybrid of trophoblastic origin, providing a unique cell culture model to study trophoblast-related invasion and its underlying genetic mechanisms.
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ABSTRACT: The differential reaction of cytotrophoblast and syncytiotrophoblast towards infection with listeria monocytogenes (LM) is pivotal to its pathogenicity. In this study we tested the cytokine signature upon infection with listeria monocytogenes (LM) in an in vitro model. We compared two related trophoblastic cell lines (AC-1M32 and ACH1P). The cell line ACH1P showed syncytium formation, whereas AC-1M32 did not fuse, as demonstrated with immunfluorescence E-Cadherin staining. In a Multi-Analyte ELISArray we tested for concentrations of TNFα, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-17A, IL-8, MCP1, MIP-1a, MIP-1b, MDC, Eotaxin, IFNγ, G-CSF, TGFβ1 after 8 and 24 hours. Compared to unstimulated cells, the syncytial cell line ACH1P showed a significant induction of Interleukin-6 (IL-6), Monocyte Chemotactic Protein-1 (MCP-1) and transforming growth factor β-1 (TGFβ1). Incubating AC-1M32 with LM, however, showed significantly reduced IL-6 levels and a massively increased (~300fold) TGFβ1 secretion compared to unstimulated controls. A functional anti-LM immune response was only induced by the syncytium-forming cell line ACH1P. Using the two sister cell lines AC-1M32 and ACH1P in an in vitro LM-stimulation assay might facilitate research in the area of placental listeria infection.American Journal Of Reproductive Immunology 08/2012; 68(5):387-91. · 3.32 Impact Factor
- Journal of The American College of Cardiology - J AMER COLL CARDIOL. 01/2011; 57(14).
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ABSTRACT: Preeclampsia is one of the leading causes for maternal and fetal morbidity. Placental protein 13 (PP13) is a placenta specific protein and with its decreased maternal serum levels in the first trimester it is one of the most promising markers to predict the syndrome in early pregnancy. In clinical trials attempts to prevent preeclampsia have already been made using low-dose aspirin, low-molecular-weight heparin, and antioxidants such as vitamins C and E. Here we investigated the effect of these agents on PP13 and beta-hCG levels using choriocarcinoma cell lines as surrogates for primary villous trophoblast. Five different cell lines were triggered with forskolin and cultured for 48 h. Amongst the five tested cell lines BeWo cells showed the strongest increase in PP13 mRNA after forskolin treatment compared to controls. Hence these cells were used to investigate the effect of varying concentrations of vitamin C, acetylsalicylic acid (ASA), Trolox) and heparin on cell fusion and PP13 and beta-hCG levels. The response to vitamin C was a dose-dependent increase in protein expression, while the other drugs showed only modest effects. Since first trimester PP13 has been shown to be significantly decreased in women subsequently developing preeclampsia, this data might point to a beneficial effect of very early vitamin C treatment of such women already in the early first trimester of pregnancy.Placenta 03/2010; 31(5):431-8. · 3.12 Impact Factor