Cell Culture Models of Human Trophoblast—Primary Culture of Trophoblast—A Workshop Report

Frank, Hans-Georg; Genbačev, Olga; Blaschitz, Astrid; Chen, C.-P.; Clarson, LH; Évain-Brion, D.; Gardner, Lucy; Malek, Abdel; Morrish, Donald W.; Loke, Y.W.; Tarrade, Anne

doi:10.1053/plac.1999.0528

Cited by 59 publications

(41 citation statements)

References 6 publications

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“…These protocols yield to an enriched preparation of CK7-positive cells, i.e., trophoblasts, and a workshop about primary culture of human trophoblasts recommended using preparations with 50% or more CK7 positive cells (30). We routinely obtained more than 90% of CK7 positive cells with little contamination by macrophages and mesenchymal cells.…”

Section: Discussionmentioning

confidence: 99%

The PDE4 Inhibitor Rolipram Prevents NF-κB Binding Activity and Proinflammatory Cytokine Release in Human Chorionic Cells

Hervé

Schmitz

Évain-Brion

et al. 2008

The Journal of Immunology

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Spontaneous preterm delivery is linked to intrauterine inflammation. Fetal membranes are involved in the inflammatory process as an important source of mediators, and the chorion leave produces high levels of the proinflammatory cytokine TNF-α when stimulated by LPS. The transcription factor NF-κB is the main regulator of this inflammatory process and controls the production of cytokines by the chorion leave. Phosphodiesterase 4 inhibitors are recognized for their anti-inflammatory and myorelaxant effects. The purpose of this study was to investigate whether PDE4 inhibition affects the LPS signaling in human cultured chorionic cells. We showed that these cells express TLR4, the main LPS receptor, and exhibit a predominant PDE4 activity. Upon LPS challenge, PDE4 activity increases concomitantly to the induction of the specific isoform PDE4B2 and chorionic cells secrete TNF-α. LPS induces the nuclear translocation of the NF-κB p65 subunit and the activation of three different NF-κB complexes in chorionic cells. The presence of the PDE4 inhibitor rolipram reduces the TNF-α production and the activation of the three NF-κB complexes. These data indicate that the PDE4 family interacts with the LPS signaling pathway during the inflammatory response of chorionic cells. PDE4 selective inhibitors may thus represent a new therapeutic approach in the management of inflammation-induced preterm delivery.

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Section: Discussionmentioning

confidence: 99%

The PDE4 Inhibitor Rolipram Prevents NF-κB Binding Activity and Proinflammatory Cytokine Release in Human Chorionic Cells

Hervé

Schmitz

Évain-Brion

et al. 2008

The Journal of Immunology

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“…Primary human trophoblast cells, in contrast, better represent the behavior of normal human trophoblasts in vivo [23]. They have been used very effectively to elucidate key aspects of syncytiotrophoblast formation [24].…”

Section: Models To Study Trophoblast Developmentmentioning

confidence: 99%

Human Embryonic Stem Cells as Models for Trophoblast Differentiation

et al. 2008

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Trophectoderm is specified from pluripotent blastomeres at some time prior to blastocyst formation. Proliferating cytotrophoblast derived from trophectoderm is the forerunner of the entire trophoblast component of the mature human placenta, including extra-villous cytotrophoblast and syncytiotrophoblast. Recently human embryonic stem cells (hESC) have been employed to study these events in an in vitro situation. Here we review some of the work in this emerging area of trophoblast biology. We concentrate primarily on a model in which colonies of hESC are exposed to BMP4 in stem cell growth medium lacking FGF2. Under both low (4 %) and high (20 %) O 2 conditions, differentiation proceeds unidirectionally towards trophoblast from the outside of the colonies inwards, with progression fastest under high O 2 . Immunohistochemical observations performed on whole colonies combined with microarray analysis of mRNA can be employed to track developmental transitions as they occur over time and in two-dimensional space as the cells respond to BMP4.

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“…After rinsing the slides in buffer for 5 min, slides were incubated with the commercial primary antibodies (see Table 2) polyclonal goat anti-N-terminal netrin-1 (dilution 1:50), rabbit anti-C-terminal netrin-1 (dilution 1:50), goat anti-UNC5B (dilution 1:50) for 30 min, or monoclonal mouse antihuman DCC antibody (BD Pharmingen; San Diego, CA, diluted 1:100) for 40 min at 37C. Mouse monoclonal anti-cytokeratin 7 (Ventana) was used for 28 min at room temperature for villous and extravillous cytotrophoblast characterization (Frank et al 2001). For extravillous cell characterization, rabbit monoclonal anti-HER-2/neu (anti-c-ErbB2; Ventana) was used for 16 min.…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…Immunoreactivity to cytokeratin 7 and c-ErbB2 antibodies was used to identify villous cytotrophoblasts, syncytiotrophoblasts, and extravillous cytotrophoblasts. Cytokeratin 7 staining identifies all villous and extravillous cytotrophoblasts (Frank et al 2000), whereas the proto-oncogene product c-ErbB2 is present in the extravillous cells (Jokhi et al 1994). Anti-vimentin antibodies were used to detect stromal cells (Kelley et al 1990).…”

Section: Localization Of Netrin-1 Unc5b and DCC In Placenta By Immumentioning

confidence: 99%

Characterization and Expression of Netrin-1 and Its Receptors UNC5B and DCC in Human Placenta

Dakouane-Giudicelli

Duboucher

Fortemps

et al. 2009

J Histochem Cytochem.

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S U M M A R Y Netrins are a family of proteins that mediate axonal guidance in the central nervous system (CNS). In addition to the CNS, netrins are involved in cell adhesion, motility, proliferation, differentiation, and survival. Because these processes occur in the placenta, we raised the question of whether netrin-1 is expressed by placental cells during development.In the present study, we analyzed the spatial and temporal distribution of netrin-1 and its two receptors, DCC (deleted in colorectal cancer) and UNC5B (uncoordinated-5 homolog) in human placenta using RT-PCR, Western blotting, and immunohistochemistry analysis. We demonstrated the presence of the proteins and transcripts of netrin-1 and its receptors in placenta and cytotrophoblasts. Furthermore, using immunohistochemistry, we localized endogenous netrin-1 protein staining to villous and extravillous cytotrophoblasts, and secreted netrin-1 outside the syncytiotrophoblasts. The DCC receptor was localized to syncytiotrophoblasts and invasive extravillous cytotrophoblasts during the first trimester and at term. On the other hand, the UNC5B receptor was localized to villous and extravillous cytotrophoblasts proximal to anchoring areas during the first trimester. At term, UNC5B was observed in decidual cells and weakly in extravillous cells. The discrete pattern of netrin-1 and netrin-1 receptor distribution suggested that netrin-1 protein functions might vary with its localization in the placenta and probably with time of gestation. (J Histochem Cytochem 58:73-82, 2010)

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Cell Culture Models of Human Trophoblast—Primary Culture of Trophoblast—A Workshop Report

Cited by 59 publications

References 6 publications

The PDE4 Inhibitor Rolipram Prevents NF-κB Binding Activity and Proinflammatory Cytokine Release in Human Chorionic Cells

The PDE4 Inhibitor Rolipram Prevents NF-κB Binding Activity and Proinflammatory Cytokine Release in Human Chorionic Cells

Human Embryonic Stem Cells as Models for Trophoblast Differentiation

Characterization and Expression of Netrin-1 and Its Receptors UNC5B and DCC in Human Placenta

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