Binding of Human IgG and F(ab′)2and Fc Fragments to Cultured Trophoblast Cells from Human Term Placenta

Ugele, Bernhard; Hecht, Joachim; Kuß, E.

doi:10.1006/excr.1998.4161

Cited by 11 publications

(9 citation statements)

References 49 publications

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“…Mononuclear cytotrophoblasts were purified from term placentae as described (18,19). The purity of the cell preparation was routinely assessed to be 90%, as determined by immunological staining for vimentin and cytokeratin as described (19).…”

Section: Isolation Of Cytotrophoblastsmentioning

confidence: 99%

“…The purity of the cell preparation was routinely assessed to be 90%, as determined by immunological staining for vimentin and cytokeratin as described (19). Cytotrophoblasts were plated at a density of 1.25 ϫ 10 5 cells/cm 2 in medium 199 supplemented with 20% FCS.…”

Section: Isolation Of Cytotrophoblastsmentioning

confidence: 99%

“…It relies on the delivery of precursors primarily from the fetal compartment, with some contribution from maternal sources, and on biosynthetic pathways abundant in the trophoblasts (1). The C 19 steroid, dehydroepiandrosterone (DHEA) and its sulfated conjugate (DHEAS), are synthesized in the fetal adrenal cortex from pregnenolone and 17-OH pregnenolone, reactions mediated by P450c17. DHEAS is in large part converted to 16-hydroxy-DHEAS exclusively in the fetal liver, and these steroid precursor can reach fetal arterial concentrations of 4 M and 10 m, respectively (4).…”

mentioning

confidence: 99%

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Characterization of an Organic Anion-Transporting Polypeptide (OATP-B) in Human Placenta

St‐Pierre

Hagenbuch

Ugele

et al. 2002

The Journal of Clinical Endocrinology &Amp; Metabolism

112

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Organic anion-transporting polypeptides (OATPs) are a family of multispecific carriers that mediate the sodium-independent transport of steroid hormone and conjugates, drugs, and numerous anionic endogenous substrates. We investigated whether members of the OATP gene family could mediate fetal-maternal transfer of anionic steroid conjugates in the human placenta. OATP-B (gene symbol SLC21A9) was isolated from a placenta cDNA library. An antiserum to OATP-B detected an 85-kDa protein in basal but not apical syncytiotrophoblast membranes. Immunohistochemistry of first-, second-, and third-trimester placenta showed staining in the cytotrophoblast membranes and at the basal surface of the syncytiotrophoblast. Trophoblasts that reacted with an antibody to Ki-67, a proliferation-associated antigen, expressed lower levels of OATP-B. OATP-B mRNA levels were measured in isolated trophoblasts under culture conditions that promoted syncytia formation. Real-time quantitative PCR estimated an 8-fold increase in OATP-B expression on differentiation to syncytia. The uptake of [ 3 H]estrone-3-sulfate, a substrate for OATP-B, was measured in basal syncytiotrophoblast membrane vesicles. Transport was saturable and partially inhibited by pregnenolone sulfate, a progesterone precursor. Pregnenolone sulfate also partially inhibited OATP-B-mediated transport of estrone-3-sulfate in an oocyte expression system. These findings suggest a physiological role for OATP-B in the placental uptake of fetal-derived sulfated

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Section: Isolation Of Cytotrophoblastsmentioning

confidence: 99%

Section: Isolation Of Cytotrophoblastsmentioning

confidence: 99%

mentioning

confidence: 99%

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Characterization of an Organic Anion-Transporting Polypeptide (OATP-B) in Human Placenta

St‐Pierre

Hagenbuch

Ugele

et al. 2002

The Journal of Clinical Endocrinology &Amp; Metabolism

112

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“…The mononucleated cytotrophoblasts did not proliferate, but differentiated within 90 h in culture in serum containing medium to multinucleated, syncytiotrophoblast-like cells (Kliman et al, 1986;Ugele et al, 1998). During this process, development of STS, PALP, and G6PD was examined by immunocytochemical techniques and by measurements of enzyme activity.…”

confidence: 99%

“…The viability, as assessed by trypan blue exclusion, exceeded 95 %. Purification and identity of isolated cells were assessed by immunocytochemical staining and release of human chorionic gonadotropin (Ugele et al, 1998). Usually, more than 90 % of the cells did not stain for vimentin but did stain for cytokeratin, indicating the trophoblast cell type (Douglas and King, 1989).…”

Section: Preparation and Cultivation Of Mononuclear Cellsmentioning

confidence: 99%

Differential increase of steroid sulfatase activity in XX and XY trophoblast cells from human term placenta with syncytia formation in vitro

Ugele¹,

Regemann²

2000

Cytogenet Genome Res

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Steroid sulfatase (STS, EC 3.1.6.2) catalyzes the hydrolysis of the sulfate ester bonds of a variety of sulfated steroids, such as cholesterol, dehydroepiandrosterone, and estrone sulfate, a reaction influencing fertility and breast cancer in mammals. The activity of the enzyme is substantially elevated in placental syncytiotrophoblasts and is lower in other somatic cells. The polypeptide sequence of the enzyme is encoded by a gene located on the distal short arm of the human X chromosome. Prior studies have shown that the STS gene escapes X-chromosome inactivation. We studied the expression of the STS gene in primary cultures of cytotrophoblasts from human term placentae and compared it with the expression of autosomally encoded placental alkaline phosphatase (PALP) and X-linked glucose-6-phosphate dehydrogenase (G6PD). During 90 h in culture, the mononucleated cytotrophoblast cells did not proliferate, but differentiated into multinucleated, syncytiotrophoblast-like cells. STS activity in freshly isolated cytotrophoblasts was low (about 17%), compared to placental tis- sue, and about 1.7-fold higher in female than in male cells. During cultivation, STS activity increased 2- to 3-fold in female, but not in male, cells. PALP activity was very low in freshly isolated cytotrophoblasts (about 5% of placental tissue), and no significant difference between female and male cells was detectable. Within 90 h in culture, PALP activity increased in all preparations about 2- to 4-fold. G6PD activity in freshly isolated cytotrophoblasts showed great variation among preparations, and no significant difference between female and male cells was detectable. In both male and female cells the activity declined to about 50% of initial activity during cultivation. We conclude that human cytotrophoblasts in primary culture show a sex-specific regulation of STS activity, perhaps as a unique feature of the STS gene. The cytotrophoblast system may offer a new possibility to study the regulation of STS gene expression.

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Hepatitis C virus infection of human cytotrophoblasts cultured in vitro

Nie

Gao

Cheng

et al. 2012

Journal of Medical Virology

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Hepatitis C virus (HCV) infection in the uterus is a significant path of vertical HCV transmission. Some studies consider vertical HCV transmission in the uterus as the result of maternal blood leakage into infant blood, whereas others theorize that HCV is transmitted by the mother to the infant through cells constituting the placenta barrier. Although trophoblasts play an important role in the placenta barrier, no definitive evidence has been presented to prove that cytotrophoblasts can be infected with HCV. The current study investigated whether or not these can be infected with HCV by conducting an experiment, in which cultured human cytotrophoblasts were infected with HCV in vitro. The results were analyzed using reverse transcription polymerase chain reaction (RT-PCR), ultrastructural characteristic changes under an electron microscope, and immunoelectron microscopy. HCV RNA in the supernatant of the cultured medium of the infected group was intermittently detected during the 16-day incubation period using RT-PCR. Under an electron microscope, the ultrastructures of infected human cytotrophoblasts were markedly different from normal cells, demonstrating lysosomal hyperplasia, rough endoplasmic reticulum, decreased lipid droplets, presence of vacuoles, and the appearance of HCV-like particles. Using immunoelectron microscopy, HCV-like particles conjoined with golden granules were also observed. Based on the data, the current study concludes that HCV infects a human cytotrophoblast cultured in vitro; moreover, its ultrastructure changes dramatically upon infection.

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Binding of Human IgG and F(ab′)2and Fc Fragments to Cultured Trophoblast Cells from Human Term Placenta

Cited by 11 publications

References 49 publications

Characterization of an Organic Anion-Transporting Polypeptide (OATP-B) in Human Placenta

Characterization of an Organic Anion-Transporting Polypeptide (OATP-B) in Human Placenta

Differential increase of steroid sulfatase activity in XX and XY trophoblast cells from human term placenta with syncytia formation in vitro

Hepatitis C virus infection of human cytotrophoblasts cultured in vitro

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