Trophoblast Cell Line

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Yasufumi Sawada - One of the best experts on this subject based on the ideXlab platform.

  • H(+)-linked transport of salicylic acid, an NSAID, in the human Trophoblast Cell Line BeWo.
    American Journal of Physiology-cell Physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and l-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intracel...

  • h linked transport of salicylic acid an nsaid in the human Trophoblast Cell Line bewo
    American Journal of Physiology-cell Physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and l-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intracel...

  • H(+)-linked transport of salicylic acid, an NSAID, in the human Trophoblast Cell Line BeWo.
    American journal of physiology. Cell physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and L-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intraCellular pH (pH(i)). The uptakes of [(14)C]salicylic acid and L-[(14)C]lactic acid were temperature- and extraCellular pH-dependent and saturable at higher concentrations. Both uptakes were also reduced by FCCP, nigericin, and NaN(3). Various nonsteroidal anti-inflammatory drugs (NSAIDs) strongly inhibited the uptake of L-[(14)C]lactic acid. Salicylic acid and ibuprofen noncompetitively inhibited the uptake of L-[(14)C]lactic acid. alpha-Cyano-4-hydroxycinnamate (CHC), a monocarboxylate transporter inhibitor, suppressed the uptake of L-[(14)C]lactic acid but not that of [(14)C]salicylic acid. CHC also suppressed the decrease of pH(i) induced by L-lactic acid but had little effect on that induced by salicylic acid or diclofenac. These results suggest that NSAIDs are potent inhibitors of lactate transporters, although they are transported mainly by a transport system distinct from that for L-lactic acid.

Gil Mor - One of the best experts on this subject based on the ideXlab platform.

  • Establishment and characterization of a new human first trimester Trophoblast Cell Line, AL07
    Placenta, 2020
    Co-Authors: Hong Liu, Liling Wang, Yan Wang, Qian Zhu, Paulomi Aldo, Jiahui Ding, Gil Mor, Ai-hua Liao
    Abstract:

    Abstract IntroductionThe limited Cell number of primary Trophoblasts and contamination of Trophoblast Cell Lines promote us to develop a novel stable Trophoblast Cell Line. Method of study; Primary Trophoblast Cells were isolated from first-trimester placenta and telomerase-induced immortalization was used to immortalize these Cells. Subsets of Cells were then evaluated by flow cytometry using CK7, HLA-G, CD45 and CD14, specific markers for Trophoblast Cells, extra-villous Trophoblast, pan leucocyte and monocyte/macrophage, respectively. Immunofluorescence staining and immunocytochemistry were used to detect CK7 expression in Trophoblast Cells. The level of secreted human Chorionic Gonadotropin (hCG) was measured by electrochemiluminescence (ECL). The Bio-Plex MAGPIX System was used to analyze the cytokines and chemokines produced by AL07 Cell Line. ResultsWe were able to isolate primary Trophoblast Cells from several first-trimester placentas. One clone, AL07 Trophoblast Cells, isolated from a week 7 placenta, was morphologically stable and positive for the expression of CK7 by immunofluorescence and immunocytochemistry staining. Characterization of AL07 Cells reveled that they are CD45 or CD14 negative and had constitutive secretion of hCG and low HLA-G expression. Furthermore, clone AL07 secret high levels of several cytokines and chemokines, including IL-6, IL-8 and VEGF, and moderately secreted MCP-1 IP-10 and RANTES. DiscussionWe report the successful isolation, immortalization and characterization of AL07 Cells, a novel Cell clone isolated from first trimester human placenta. The clone is free of contamination of immune Cells, and exhibits similar cytokine profile as other Trophoblast Cell Lines. This new cytoTrophoblast-like AL07 Cell, can be a valuable tool for in-vitro Trophoblast studies in the future.

  • Cytogenetic features of human Trophoblast Cell Lines SWAN-71 and 3A-subE.
    Placenta, 2017
    Co-Authors: Jill L. Reiter, Holli M. Drendel, Sujata Chakraborty, Megan M. Schellinger, Men-jean Lee, Gil Mor
    Abstract:

    Immortalization of primary Cells with telomerase is thought to maintain normal phenotypic properties and avoid chromosomal abnormalities and other cancer-associated changes that occur following simian virus 40 tumor antigen (SV40 Tag) induced immortalization. However, we report that the human telomerase reverse transcriptase (hTERT)-immortalized SWAN-71 Trophoblast Cell Line has a near pentaploid 103∼119,XXXX[cp20] karyotype. Additionally, DNA typing analysis indicated that SWAN-71 Cells have acquired microsatellite instability. In comparison, the post-crisis SV40-transformed Trophoblast Cell Line 3A-subE was hypertriploid 69∼81,XX[cp20]. Both Cell Lines contained multiple specific clonal rearrangements. These findings emphasize the need to monitor for genetic instability in hTERT-immortalized Cells.

  • the isolation and characterization of a novel telomerase immortalized first trimester Trophoblast Cell Line swan 71
    Placenta, 2009
    Co-Authors: Shawn L Straszewskichavez, Roberto Romero, Seth Guller, Paulomi Aldo, Vikki M Abrahams, Ayesha B Alvero, Gil Mor
    Abstract:

    Studies using first trimester Trophoblast Cells may be limited by the inability to obtain patient samples and/or adequate Cell numbers. First trimester Trophoblast Cell Lines have been generated by SV40 transformation or similar methods, however, this approach is known to induce phenotypic and karyotypic abnormalities. The introduction of telomerase has been proposed to be a viable alternative for the immortalization of primary human Cells. To investigate whether telomerase-induced immortalization might be a more feasible approach for the generation of first trimester Trophoblast Cell Lines, we isolated primary Trophoblast Cells from a 7-week normal placenta and infected the Cells with human telomerase reverse transcriptase (hTERT), the catalytic subunit of telomerase. Although this hTERT-infected first trimester Trophoblast Cell Line, which we have named Swan 71, has been propagated for more than 100 passages, it still has attributes that are characteristic of primary first trimester Trophoblast Cells. The Swan 71 Cells are positive for the expression of cytokeratin 7, vimentin and HLA-G, but do not express CD45, CD68 or the Fibroblast Specific Antigen (FSA), CD90/Thy-1. In addition, we also demonstrated that the Swan 71 Cells secrete fetal fibronectin (FFN) as well as low levels of human Chorionic Gonadotrophin (hCG). Moreover, the Swan 71 Cells exhibit a cytokine and growth factor profile that is similar to primary Trophoblast Cells and are resistant to Fas, but not TNF-alpha-induced apoptosis. This suggests that the Swan 71 Cells may represent a valuable model for future in vitro Trophoblast studies.

  • A potential tolerogenic immune mechanism in a Trophoblast Cell Line through the activation of chemokine-induced T Cell death and regulatory T Cell modulation
    Human reproduction (Oxford England), 2008
    Co-Authors: Laura Fraccaroli, Gil Mor, Julio Armando Alfieri, Luciana Larocca, Mario Jose Calafat, Claudia Pérez Leirós, Rosanna Ramhorst
    Abstract:

    BACKGROUND Successful implantation is followed by a local pro-inflammatory and Th1 response, subsequently controlled by Th2. Regulated upon activation, normal T Cell expressed and secreted (RANTES) promotes a Th1 response and is implicated as a physiologic tolerogenic factor; therefore, we studied its potential role in the Trophoblast-maternal leukocyte dialog. METHODS We performed co-cultures of immortalized Trophoblast Cell Line (Swan 71) and peripheral blood mononuclear Cells (PBMCs) from fertile women (n = 23) or with recurrent spontaneous abortions (n = 18, RSA). After 24 and 48 h, supernatant and Cells were analyzed by enzyme-linked immunosorbent assay, fluorescence-activated Cell sorting, Western blot and apoptosis assay. To investigate the physiological effects at peripheral level, we co-cultured maternal and paternal PBMCs with conditioned media from Swan Cells and progesterone. RESULTS Following interaction of maternal PBMCs and Trophoblast Cells, RANTES production increased (P < 0.05) and was accompanied by low levels of interferon gamma, interleukin-12 p70 and high levels of tumor necrosis factor-alpha, nitrites and leukemia-inhibitory factor. RANTES production resulted in elevated apoptosis of potentially deleterious maternal CD3+ lymphocytes, accompanied by a decrease in the proliferative maternal response. During fetal-maternal dialog, the anti-RANTES antibody significantly reduced the frequency of CD4+CD25+Foxp3+ Cells (P < 0.05) and was associated with Trophoblast Cell survival. However, co-cultures of Swan Cells and RSA-PBMCs displayed a differential RANTES kinetics, lower levels of regulatory T Cells (Tregs) and CD3+annexin-V+Cells, accompanied by higher levels of apoptotic Trophoblast Cells. CONCLUSIONS RANTES promotes an adequate pro-implantatory microenvironment that influences Trophoblast Cell survival and modulates the balance of maternal Treg/T effector lymphocytes in favor of maternal tolerance.

Andy J.g. Pötgens - One of the best experts on this subject based on the ideXlab platform.

Akiko Emoto - One of the best experts on this subject based on the ideXlab platform.

  • H(+)-linked transport of salicylic acid, an NSAID, in the human Trophoblast Cell Line BeWo.
    American Journal of Physiology-cell Physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and l-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intracel...

  • h linked transport of salicylic acid an nsaid in the human Trophoblast Cell Line bewo
    American Journal of Physiology-cell Physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and l-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intracel...

  • H(+)-linked transport of salicylic acid, an NSAID, in the human Trophoblast Cell Line BeWo.
    American journal of physiology. Cell physiology, 2002
    Co-Authors: Akiko Emoto, Fumihiko Ushigome, Noriko Koyabu, Hiroshi Kajiya, Koji Okabe, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Hisakazu Ohtani, Yasufumi Sawada
    Abstract:

    We investigated the transport of salicylic acid and L-lactic acid across the placenta using the human Trophoblast Cell Line BeWo. We performed uptake experiments and measured the change in intraCellular pH (pH(i)). The uptakes of [(14)C]salicylic acid and L-[(14)C]lactic acid were temperature- and extraCellular pH-dependent and saturable at higher concentrations. Both uptakes were also reduced by FCCP, nigericin, and NaN(3). Various nonsteroidal anti-inflammatory drugs (NSAIDs) strongly inhibited the uptake of L-[(14)C]lactic acid. Salicylic acid and ibuprofen noncompetitively inhibited the uptake of L-[(14)C]lactic acid. alpha-Cyano-4-hydroxycinnamate (CHC), a monocarboxylate transporter inhibitor, suppressed the uptake of L-[(14)C]lactic acid but not that of [(14)C]salicylic acid. CHC also suppressed the decrease of pH(i) induced by L-lactic acid but had little effect on that induced by salicylic acid or diclofenac. These results suggest that NSAIDs are potent inhibitors of lactate transporters, although they are transported mainly by a transport system distinct from that for L-lactic acid.

Fátima Martel - One of the best experts on this subject based on the ideXlab platform.

  • Xanthohumol impairs glucose uptake by a human first-trimester extravillous Trophoblast Cell Line (HTR-8/SVneo Cells) and impacts the process of placentation
    Molecular human reproduction, 2015
    Co-Authors: Ana Correia-branco, Elisa Keating, Cláudia Azevedo, João R. Araújo, João Tiago Guimarães, Ana Faria, Fátima Martel
    Abstract:

    In this study, we aimed to investigate modulation of glucose uptake by the HTR-8/SVneo human first-trimester extravillous Trophoblast Cell Line by a series of compounds and to study its consequences upon Cell proliferation, viability and migration. We observed that uptake of (3)H-deoxy-d-glucose ((3)H-DG; 10 nM) was time-dependent, saturable, inhibited by cytochalasin B (50 and 100 µM), phloretin (0.5 mM) and phloridzin (1 mM), insulin-insensitive and sodium-independent. In the short term (30 min), neither 5-HT (100-1000 µM), melatonin (10 nM) nor the drugs of abuse ethanol (100 mM), nicotine (100 µM), cocaine (25 µM), amphetamine (10-25 µM) and 3,4-methylenedioxy-N-methamphetamine (10 µM) affected (3)H-DG uptake, while dexamethasone (100-1000 µM), fluoxetine (100-300 µM), quercetin, epigallocatechin-3-gallate (30-1000 µM), xanthohumol (XH) and resveratrol (1-500 µM) decreased it. XH was the most potent inhibitor [IC50 = 3.55 (1.37-9.20) µM] of (3)H-DG uptake, behaving as a non-competitive inhibitor of (3)H-DG uptake, both after short- and long-term (24 h) treatment. The effect of XH (5 µM; 24 h) upon (3)H-DG uptake involved mammalian target of rapamycin, tyrosine kinases and c-Jun N-terminal kinases intraCellular pathways. Moreover, XH appeared to decrease Cellular uptake of lactate due to inhibition of the monocarboxylate transporter 1. Additionally, XH (24 h; 5 µM) decreased Cell viability, proliferation, culture growth and migration. The effects of XH upon Cell viability and culture growth, but not the antimigratory effect, were mimicked by low extraCellular glucose conditions and reversed by high extraCellular glucose conditions. We thus suggest that XH, by inhibiting glucose Cellular uptake and impairing HTR-8/SVneo Cell viability and proliferation, may have a deleterious impact in the process of placentation.

  • Acute and chronic effects of some dietary bioactive compounds on folic acid uptake and on the expression of folic acid transporters by the human Trophoblast Cell Line BeWo.
    The Journal of nutritional biochemistry, 2007
    Co-Authors: Elisa Keating, Clara Lemos, Pedro Gonçalves, Fátima Martel
    Abstract:

    Folic acid (FA) is a vitamin that acts as a coenzyme in the biosynthesis of purine and pyrimidine precursors of nucleic acids, which are critically important during pregnancy. Our group has previously shown that both reduced folate carrier (RFC1) and folate receptor alpha (FRalpha) seem to be involved in the uptake of [3H]folic acid ([3H]FA) by a human Trophoblast Cell Line (BeWo) and by human primary cultured cytoTrophoblasts. Our aim was to study the interaction between FA and some nutrients/bioactive substances. For this, we tested the acute and chronic effects of some dietary compounds on [3H]FA apical uptake and on the expression of both RFC1 and FRalpha mRNA in BeWo Cells. Our results show that [3H]FA uptake was significantly reduced by acute exposure to epicatechin, isoxanthohumol (1-400 microM) or theophylLine (0.1-100 microM); isoxanthohumol seemed to act as a competitive inhibitor, whereas epicatechin and theophylLine caused an increase in both Km and Vmax. On the other hand, [3H]FA uptake was significantly increased by chronic exposure to xanthohumol, quercetin or isoxanthohumol (0.1-10 microM), and this increase does not seem to result from changes in the level of RFC1 or FRalpha gene expression. Moreover, [3H]FA uptake was significantly reduced by chronic exposure to ethanol (0.01%). This reduction seems to be, at least in part, due to a reduction in FRalpha expression. These results are compatible with an association between a deficient FA supply to the placenta/fetus and ethanol toxicity in pregnancy.

  • Characteristics of thiamine uptake by the BeWo human Trophoblast Cell Line.
    Journal of Biochemistry and Molecular Biology, 2006
    Co-Authors: Elisa Keating, Clara Lemos, Isabel Azevedo, Fátima Martel
    Abstract:

    Little is known concerning the mechanisms responsible for the transplacental transfer of thiamine. So, the aim of this work was to characterize the placental uptake of thiamine from the maternal circulation, by determining the characteristics of -thiamine uptake by a human Trophoblast Cell Line (BeWo). Uptake of -thiamine (50-100 nM) by BeWo Cells was: 1) temperature-dependent and energy-independent; 2) pH-dependent (uptake increased as the extraCellular medium pH decreased); 3) -dependent and -independent; 4) not inhibited by the thiamine structural analogs amprolium, oxythiamine and thiamine pyrophosphate; 5) inhibited by the unrelated organic cations guanidine, N-methylnicotinamide, tetraethylammonium, clonidine and cimetidine; 6) inhibited by the organic cation serotonin, and by two selective inhibitors of the serotonin plasmalemmal transporter (hSERT), fluoxetine and desipramine. We conclude that -thiamine uptake by BeWo Cells seems to occur through a process distinct from thiamine transporter-1 (hThTr-1) and thiamine transporter-2 (hThTr-2). Rather, it seems to involve hSERT. Moreover, chronic (48 h) exposure of Cells to caffeine () stimulated and chronic exposure to xanthohumol and iso-xanthohumol (1 and , respectively) inhibited -thiamine uptake, these effects being not mediated through modulation of the expression levels of either hThTr-1 or hSERT mRNA.