Differentiation of Human Trophoblasts: Structure-Function Relationships

Kao, Lee-Chuan; Babalola, Gbolagade O.; Kopf, Gregory S.; Coutifaris, Christos; StraussIII, Jerome F.

doi:10.1007/978-1-4613-9260-6_10

Cited by 10 publications

(8 citation statements)

References 23 publications

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“…The data presented as well as the previously reported time course of endogenous progesterone secretion in this cell model of trophoblast differentiation (14,17,19) support these hypotheses. Using the present in vitro model of trophoblast differentiation, we have previously shown that trophoblast aggregation is associated with an increase in intracellular cAMP (12), and thus, the observation of agonist activity when the cells are exposed to RU486 alone at a time of low endogenous progesterone (24h) agrees with results obtained using other experimental models. Nevertheless, as pro-gesterone receptors have been shown to be present in cytotrophoblasts (20), an effect of progesterone at the level of the placenta should also be considered.…”

Section: Discussionsupporting

confidence: 88%

“…The results presented further extend our understanding of how the expression of this secretory protein is regulated and provide insight into its potential role in the human placenta. Both in vivo and in vitro, the morphological differentiation of these cells is accompanied by functional differentiation with respect to hormone production, including progesterone (12,14,18). In vivo, these cells exist in an endocrine milieu that is high in progesterone produced by the overlying syncytiotrophoblast layer of the chorionic villus, and we hypothesize that this contributes to the high levels of OPN mRNA expressed by freshly isolated cytotrophoblasts.…”

Section: Discussionmentioning

confidence: 90%

“…The membrane was prehybridized with 50% (vol/vol) formamide, standard saline phosphate, and ethylenediamine acetate buffer (SSPE), 1 ϫ Denhardt's solution, and 0.1% SDS at 42 C for 2 h. A 1493-bp complementary DNA (cDNA) encoding the entire protein region of human OPN (a gift from M. Young, NIH) served as the template for synthesis of a labeled DNA using random priming (12) with [ 32 P]deoxy-CTP (New England Nuclear Corp., Boston, MA), a mixture of the other deoxynucleoside triphosphates, and Klenow enzyme (Random Primed DNA Labeling Kit, Boehringer Mannheim Biochemicals, Indianapolis, IN) to a specific activity of 1 ϫ 10 6 to 1 ϫ 10 7 cpm/ng probe. The membrane was prehybridized with 50% (vol/vol) formamide, standard saline phosphate, and ethylenediamine acetate buffer (SSPE), 1 ϫ Denhardt's solution, and 0.1% SDS at 42 C for 2 h. A 1493-bp complementary DNA (cDNA) encoding the entire protein region of human OPN (a gift from M. Young, NIH) served as the template for synthesis of a labeled DNA using random priming (12) with [ 32 P]deoxy-CTP (New England Nuclear Corp., Boston, MA), a mixture of the other deoxynucleoside triphosphates, and Klenow enzyme (Random Primed DNA Labeling Kit, Boehringer Mannheim Biochemicals, Indianapolis, IN) to a specific activity of 1 ϫ 10 6 to 1 ϫ 10 7 cpm/ng probe.…”

Section: Rna Isolation and Northern Analysismentioning

confidence: 99%

“…These observations provided the opportunity for study of the regulation of OPN expression in a human cell model. The major hormones produced by trophoblasts, and specifically syncytiotrophoblasts, in the later stages of their differentiation in vitro are hCG and progesterone (11,12). We were prompted to perform additional experiments in which the cells were cultured for longer periods due to an observed upward trend in mRNA abundance at the later stages of the in vitro differentiation of the cells.…”

mentioning

confidence: 99%

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Progesterone Regulates Osteopontin Expression in Human Trophoblasts: A Model of Paracrine Control in the Placenta?*

et al. 1997

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Osteopontin (OPN), a matrix glycosylated phosphoprotein, has been proposed to play a role(s) in basic cellular processes, such as neovascularization and tissue remodeling, which are essential to placental morphogenesis and embryo implantation. We have shown OPN to be expressed by cytotrophoblasts of the chorionic villus, and a putative progesterone regulatory element in the OPN promoter suggests hormonal regulatory control. This led us to test the hypothesis that progesterone regulates OPN expression in human cytotrophoblasts. Cytotrophoblasts isolated from human placentas were treated with combinations of progesterone, RU486, and/or aminoglutethimide, and their expression of OPN was assessed by Northern hybridization and immunocytochemistry. The expression of OPN messenger RNA (mRNA) declined as trophoblasts aggregated, but rebounded at later times when syncytia and mononuclear cytotrophoblasts coexisted in culture. Progesterone increased OPN mRNA expression by aggregating mononuclear cytotrophoblasts. Aminoglutethimide suppression of endogenous steroidogenesis by syncytiotrophoblasts inhibited OPN expression, whereas the addition of exogenous progesterone to cells treated with aminoglutethimide reversed this inhibitory effect. These observations were confirmed at the protein level by immunocytochemistry. Treatment of cytotrophoblasts with both progesterone and RU486 inhibited the up-regulatory effect on OPN mRNA associated with exposure to progesterone alone, further confirming a direct effect of progesterone. We conclude that progesterone up-regulates OPN expression in human cytotrophoblasts, and we propose that in vivo, progesterone secretion by syncytiotrophoblasts regulates the expression of OPN by the underlying cytotrophoblasts. As the receptors for OPN, alpha(v) integrins, are expressed by syncytiotrophoblasts, we postulate that these paracrine regulatory mechanisms contribute to the adhesive and/or signaling events between the two trophoblast cell types of the chorionic villus.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 90%

Section: Rna Isolation and Northern Analysismentioning

confidence: 99%

mentioning

confidence: 99%

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Progesterone Regulates Osteopontin Expression in Human Trophoblasts: A Model of Paracrine Control in the Placenta?*

et al. 1997

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“…This enhanced the expression of genes representative of syncytiotrophoblast endocrine activity (Kao et al 1992). It is also reported that treatment with 8-bromo-cAMP causes cytotrophoblasts isolated from term placenta to aggregate within hours and an increase in hCG and progestin secretion in serum-supplemented medium (Feinman et al 1986).…”

Section: Discussionmentioning

confidence: 80%

8-bromo-cyclicAMP stimulates glucose transporter-1 expression in a human choriocarcinoma cell line

Ogura¹,

Sakata²,

Okamoto³

et al. 2000

Journal of Endocrinology

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Facilitative glucose transporter-1 (GLUT1) is abundant in trophoblast cells and is responsible for glucose transport in the placenta. However, the change in GLUT expression in human placenta upon trophoblast differentiation remains to be clarified. Therefore, we first examined the localization of GLUT1 and GLUT3 using human firsttrimester chorionic villi. We found that GLUT1 and GLUT3 were mainly localized to syncytiotrophoblast and cytotrophoblast cells respectively. We analyzed whether placental GLUT1 and GLUT3 expression changes during differentiation using a human choriocarcinoma (BeWo) cell line which is known to show functional and morphological differentiation in response to cAMP in culture. Treatment of BeWo cells with 8-bromocyclicAMP (8-bromo-cAMP) increased the level of hCG secretion and induced cell fusion leading to the formation of large syncytia. Treatment of BeWo cells with 8-bromocAMP also resulted in a significant increase in glucose uptake on days 2-3 of culture. The stimulating effect of 8-bromo-cAMP on glucose uptake was concentration dependent. Northern and immunoblot analyses revealed that the levels of mRNA and protein of GLUT1, but not of GLUT3, were significantly increased by 8-bromocAMP. These findings suggest that 8-bromo-cAMP stimulates GLUT1 expression with differentiation in BeWo cells.

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Differential Expression of Membrane and Soluble Adenylyl Cyclase Isoforms in Cytotrophoblast Cells and Syncytiotrophoblasts of Human Placenta

et al. 2003

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Differentiation of Human Trophoblasts: Structure-Function Relationships

Cited by 10 publications

References 23 publications

Progesterone Regulates Osteopontin Expression in Human Trophoblasts: A Model of Paracrine Control in the Placenta?*

Progesterone Regulates Osteopontin Expression in Human Trophoblasts: A Model of Paracrine Control in the Placenta?*

8-bromo-cyclicAMP stimulates glucose transporter-1 expression in a human choriocarcinoma cell line

Differential Expression of Membrane and Soluble Adenylyl Cyclase Isoforms in Cytotrophoblast Cells and Syncytiotrophoblasts of Human Placenta

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