Human Decidual Tissue Research Articles

Synthesis and release of endothelin-1 by human decidual cells.

We have studied whether a novel vasoconstrictor, endothelin-1 (ET-1), is synthesized by and released from human decidual cells in early pregnancy, and whether ET-1 acts directly on their own cells. It was observed that ET-1-like immunoreactivity (ET-1-LI) was released from cultured decidual, but not villous, cells, as a function of time. Reverse-phase high-pressure liquid chromatography of the conditioned media from the decidual cells revealed a major peak of ET-1-LI coeluting with standard ET-1. Phorbol myristate acetate, a protein kinase C activator, dose-dependently increased the release of ET-1-LI from the decidual cells, while a protein kinase C inhibitor, H7, significantly attenuated the stimulatory effect of phorbol myristate acetate on ET-1-LI release. Northern blot analysis demonstrated the expression of messenger RNA for prepro-ET-1 in the decidual tissue, but no such messenger RNA was observed in the villous tissue. The human decidual tissue contained a noninteracting, single class of binding sites demonstrating higher affinity for ET-1 and ET-2 than ET-3. This would be most consistent with the ETA receptor subtype. An ET-1-induced, dose-dependent accumulation of total inositol phosphates was also observed in human decidual cells prelabeled with myo-[3H]inositol. The present results demonstrate for the first time that human decidual cells in early pregnancy can synthesize and release ET-1. These cells also possess specific functional receptors for ET-1 which are coupled to phosphoinositide hydrolysis. Thus our data suggest a possible role for ET-1 in autocrine and/or paracrine function in human decidual cells.

Lectin histochemistry of the human amnio-chorionic membrane complex

The presence and distribution ofglycoconjugates within the human amnion, chorion, and decidual tissues was examined histochemically following incubation with HRP-labelled lectins. Glycoconjugates within the cytoplasm of cells as well as the extracellular matrix of the maternal decidual layer and fetal chorio-amniotic membranes reacted selectively with OFA, LTA, WGA, PSA, UEA-I, GSAI-B4, RCA-I, LFA, WA, PHA-E, and GSA-II. Conjugated lectins specific for L- fucosestained components of amnion, chorion laeve, and decidua while other lectins bound glycoconjugates within decidua and at the junction between maternal decidua and fetal chorionic tissue. These observations suggest that the amnio-chorionic membrane complex may include several histochemically distinct and specialized subpopulations of glycoconjugates distributed within cells and the extracellular matrix. Although specific functions for these glycoconjugates have not been elucidated, they may mediate transport of amniotic fluid and/or facilitate maternal recognition of the developing fetus.

Spatial distribution of active genes implicated in the regulation of insulin-like growth factor stimulatory loops in human decidual and placental tissue of first-trimester pregnancy.

We have previously shown that the insulin-like growth factor-2 (IGF-2) gene is partially coexpressed with the IGF-1 and -2 receptor genes in proliferative cytotrophoblasts of the human extraembryonic tissue. Here we show that high levels of IGF-2 gene expression are not restricted to the embryonic tissue but can also be found in the decidua compacta. The IGF-2 gene is thus expressed at high levels in the mesenchymal stroma of the decidua to establish potentially short-range communication with primarily IGF-1 receptor-positive mesenchymal stroma cells. Conversely, the glandular and surface epithelia coexpress the IGF-1 receptor and IGF-1 genes, while the IGF-2 gene is not detected above background levels. The potential control mechanisms of these cell-cell signalling pathways were investigated by the analysis of the spatial distribution of active IGF binding proteins (IGFBP) genes. The IGFBP-3 gene is coexpressed with the IGF-2 gene in proliferative cytotrophoblasts of the embryonic placenta. While active IGFBP-1 and -2 genes in our hands cannot be detected in the embryonic placenta, all three IGFBP genes are expressed in complex and overlapping patterns in the decidua compacta. The results are discussed in terms of how the various IGFBP genes may operate in different cell types to restrict IGF local stimulatory pathways.

Studies of Mammalian Implantation In Vitro. (I): Production of Laminin from Human Decidual Cell Line and Its Effects on Murine Trophoblastic Outgrowth.

TTK-1 cell line from human first trimester decidual tissue carried the surface markers such as HNK-1 (Leu 7) and HLA-DR. This cell line was strongly suspected of being human natural suppressor cell line of bone marrow or lymphoid tissue origin. In addition, the cells were found to express laminin on its cell surface, as examined by indirect immunofluorescence (IIF) test, and also secrete it into culture supernates as examined by enzyme linked immunoassay (EIA) and immunostaining. The addition of the supernates from this cell line into the murine embryo culture induced the trophoblastic outgrowth of embryos in vitro in the same manner as murine EHS sarcoma derived laminin did. This phenomenon was considered to be due to laminin in the UK-1 supernates because the effect was completely blocked by the addition of anti-laminin antibodies into the embryo culture.

Colony-Stimulating Factor-1 and c-fmsExpression in Human Endometrial Tissues and Placenta during the Menstrual Cycle and Early Pregnancy*

Colony-stimulating factor-1 (CSF-1), a growth factor produced by monocytes, macrophages, fibroblasts, and endothelial cells, has been implicated in the functional regulation and growth of the murine placenta through the presence of the CSF-1 receptor, c-fms, found in this tissue. In this study we examined the tissue levels of CSF-1 by RIA and the relative expression of CSF-1 and c-fms mRNA by Northern blot analysis in human endometrial, decidual, and placental tissues during the normal menstrual cycle and early pregnancy. All endometrial, decidual, and placental tissues demonstrated extractable immunoreactive CSF-1 and expressed the 4.0-kilobase CSF-1 mRNA species. First trimester decidual tissue expressed higher levels of CSF-1 mRNA than proliferative (3.2-fold higher; P less than 0.01) or secretory (2.4-fold higher; P less than 0.01) endometrial tissues, whereas proliferative and secretory endometrial tissues expressed similar levels of CSF-1 mRNA. Tissue extractable levels of immunoreactive CSF-1 were 3.2-fold (P less than 0.05) higher in first trimester decidual tissue and 2.9-fold (P less than 0.05) higher in secretory endometrial tissue compared to levels in proliferative endometrial tissue, whereas first trimester decidua and secretory endometrial tissues had similar levels of immunoreactive CSF-1. There was expression of c-fms mRNA in all endometrial and first trimester decidual tissue samples, with little change during the menstrual cycle and early pregnancy. In placenta, there was a positive correlation of increasing CSF-1 and c-fms mRNA expression with increasing gestational age. These results suggest that there is increased local production of CSF-1 in tissues found at the maternal-fetal interface during the time of implantation and early pregnancy. This increased production of CSF-1 may play a role in decidual function and placental growth through the presence of c-fms in these tissues.

Isolation and purification of microvascular endothelium from human decidual tissue in the late phase of pregancy

Normal pregnancy demands significant structural and physiologic adaptations of the uterine microvasculature, to facilitate adequate placentation. In pregnancies complicated by pregnancy-associated hypertension (preeclampsia), this process is defective, resulting in a high incidence of intrauterine growth retardation. The microvascular endothelium, a focal point for both initiation and inhibition of coagulation and control of vascular tone, may well contribute to development and aggravation of these abnormalities. We describe a method for isolation, purification, and culture of human decidual endothelial cells from biopsies performed at the time of cesarean section. The separation procedure is technically simple, requires only a small piece of tissue, and takes approximately 2 hours to perform. Some of the unique features of these cells in culture are outlined. This technique will permit the close examination of various aspects of the function of these cells in normal pregnancy, and their comparison with cells from pregnancies complicated by hypertension.

3. Regulation of the synthesis and release of decidual prolactin by placental and autocrine/paracrine factors

Human decidual tissue synthesizes and secretes a protein that is identical to pituitary prolactin in its chemical, biological and immunological properties. Nevertheless, the factors that regulate the synthesis and release of prolactin from the decidual tissues appear to be different to those regulating the synthesis and release of pituitary prolactin. Studies from our laboratory over the past few years indicate that the synthesis and release of decidual prolactin are regulated, at least in part, by factors released by placenta, fetal membranes and decidua. The placenta releases a 23.5 KM r protein [decidual prolactin-releasing factor (PRL-RF)] that stimulates a rapid release of prolactin within the first few minutes of exposure and a sustained, prolonged, increase in the synthesis and release of prolactin beginning 6–8 h after exposure. The acute release ofprolactin in response to PRL-RF is inhibited by decidual prolactin release-inhibitory factor (PRL-IF), a 35–45 K Mr protein that is released by the decidua. The secondary increase in the synthesis and release of prolactin in response to PRF-RF is blocked by lipocortin I, which is synthesized by both the placenta and decidua. IGF-I, insulin and relaxin also stimulate the synthesis and release of prolactin. However, the stimulation in response to these factors does not occur until 24–48 h after exposure. The cellular mechanisms involved in the release of decidual prolactin are as yet unknown. However, recent studies implicate activation of adenylate cyclase, phospholipase C-mediated phosphoinositide hydrolysis and phospholipase A 2-mediated arachidonic acid release in the regulation of prolactin release. The finding that the synthesis and release of decidual prolactin are regulated, at least in part, by PRL-RF IGF-I, insulin, relaxin and lipocortin I strongly suggests that there is novel feedback regulation between the placenta, fetal membranes, and decidua in the regulation of the synthesis and release of decidual prolactin.

The Role of Macrophages in Human Decidual Tissue during Early Spontaneous Abortion

The Role of Macrophages in Human Decidual Tissue during Early Spontaneous Abortion

Identification, immunoaffinity purification and initial characterization of a novel 71 kD human decidua associated protein by use of specific monoclonal antibodies

This study is part of an ongoing attempt to identify and characterize proteins associated with the human decidual tissue. A novel decidual-associated glycoprotein with an apparent molecular weight of 71 kD named hDP71 (human decidual-protein 71), has been identified and purified by immunoaffinity technique using monoclonal antibodies. The monoclonal antibodies recognizing the hDP71 were raised against a partly purified preparation of decidual associated proteins, which was obtained by immunoabsorption of serum proteins from crude decidual extract. Although the hDP71 was copurified with another decidual-associated glycoprotein, the previously described hDP200 (Halperin et al. (1989)), evidence is presented showing that the monoclonal antibodies described above are specific for hDP71.

The Effect of Epidermal Growth Factor (EGF) with Estradiol and Progesterone on Prolactin Secretion from Cultured Human Decidual Tissues of Early Pregnancy

Prolactin is now accepted as a normal product of the decidual cells of the human endometrium. We investigated the effect of epidermal growth factor (EGF) with estradiol and progesterone on prolactin secretion by the decidual tissues from the early pregnant endometrium. The decidual tissues were separated from villi, minced and cultured in collagen gel matrix with serum-free medium. Immunological staining of the cultured decidual tissues showed prolactin localization and EGF receptors on the stromal cells. Cultured media were collected every 2 days. The culture for the first 2 days was incubated with the serum-free medium alone (= preculture), and the following test culture was supplemented with/without additives. The prolactin content in cultured media was quantified by EIA. The results of the effect of steroid(s) and EGF were represented as a comparison of prolactin contents in the preculture and the test culture. An increase in prolactin secretion was found after the tissues were treated with a combination of 10(-8)M estradiol and 10(-6)M progesterone or 10(-6)M progesterone alone. After 8 days, the prolactin secretion rate increased about 3-fold over the precultured value. Estradiol alone kept the prolactin secretion at the precultured value. Prolactin secretion gradually decreased in the non-additive culture. These results indicate that progesterone was essential in the secretion of prolactin. Simultaneously, similar decidual tissues were incubated with a combination of EGF and steroid(s). The secretion of prolactin in the group treated with progesterone alone decreased dose-dependently responding to added EGF on the 8th day of culture. In the presence of estradiol and progesterone, the secretion rate decreased to the values similar to the progesterone alone group with the addition of 0.1, 1 ng/ml EGF, and the decrease in prolactin secretion was less with the addition of 10 ng/ml EGF. Mixed cultures of the decidual tissues and villi showed that the prolactin secretion rate increased in all groups treated with/without estradiol and/or progesterone. These results imply that progesterone derived from villi might control decidual prolactin secretion. The effect of high concentration EGF (50 ng/ml) on the prolactin secretion appeared similar to the isolated decidual tissues. These results suggest that decidual prolactin secretion is regulated by the combined effects of steroids and EGF.

The synthesis of tumour necrosis factor by human placental and decidual tissue

The synthesis of tumour necrosis factor by human placental and decidual tissue

The synthesis of TNF by human placental and decidual tissue

The synthesis of TNF by human placental and decidual tissue

Localization and Production of Human Macrophage Coloney Stimulating Factor (hM-CSF) and Granulocyte Coloney Stimulating Factor (G-CSF) in Human Placental and Decidual Tissues

Localization and Production of Human Macrophage Coloney Stimulating Factor (hM-CSF) and Granulocyte Coloney Stimulating Factor (G-CSF) in Human Placental and Decidual Tissues

Insulin-like growth factor I stimulates the synthesis and release of prolactin from human decidual cells.

Recent studies suggest a role for insulin-like growth factor I (IGF-I) in the regulation of hormone release from placental, gonadal, and pituitary tissues. To examine whether IGF-I may also regulate the release of PRL from human decidual tissue, we have investigated the effect of recombinant human IGF-I on PRL release from monolayer cultures of human decidual cells exposed to IGF-I for up to 4 days. IGF-I (10-1000 ng/ml) stimulated a sustained dose-dependent increase in PRL release (half-maximal concentration, 25 ng/ml) beginning 48 h after initial exposure, but had no effect on the intracellular PRL content. The amounts of PRL released from maximally stimulated cultures on days 3 and 4 were 168 +/- 3% (mean +/- SEM) and 258 +/- 8% of control values, respectively. IGF-I-mediated effects were inhibited by cycloheximide (3.6 microM), suggesting that the increase in PRL was the result of newly synthesized hormone. The increase in PRL release was not due to a generalized effect on protein release, since IGF-I had no effect on the release of trichloroacetic acid-precipitable [35S]methionyl proteins. Radioligand competition studies indicate that the biological actions of IGF-I are mediated through interaction with the IGF-I receptor. Binding of radiolabeled IGF-I to decidual cells in suspension was specific, saturable, and displacable by unlabeled IGF-I, with a potency nearly 10 times greater than that of insulin. Furthermore, exposure of decidual cells to a monoclonal antibody to the IGF-I receptor (alpha-IR3) completely inhibited both IGF-I-mediated PRL release and specific binding of [125I]IGF-I to decidual cells. Since the actions of IGF-I occurred at physiological concentrations, these findings strongly support a role for IGF-I in the regulation of PRL secretion by human decidua.

Decidual prolactin (PRL)-releasing factor stimulates the synthesis of PRL from human decidual cells.

Previous studies from our laboratory demonstrated that the acute release of PRL from human decidual tissue is stimulated by a 23.5 kilodalton placental protein which we designated decidual PRL-releasing factor (PRL-RF). To determine whether PRL-RF may also affect the synthesis of PRL and/or cause a secondary increase in PRL release, we have examined the effects of purified PRL-RF on the synthesis and release of PRL over a 96-h period. Exposure of dispersed decidual cells to PRL-RF (0.5 microgram/ml) stimulated a biphasic increase in PRL release with acute transient stimulation during the first 0.5 h and a delayed and sustained stimulation beginning about 8 h after exposure which persisted for the duration of the 96 h. The amounts of PRL released from PRL-RF-exposed cells after 0.5, 8, 12, 24, and 96 h were 321.2 +/- 36.2 (mean +/- SEM, n = 3), 110.2 +/- 5.3, 138.2 +/- 7.2, 194.5 +/- 11.2, and 201.5 +/- 14.2% that of control cells. Studies of the de novo synthesis of [35S]methionyl PRL indicated that the increase in PRL release after the first few hours of exposure to PRL-RF was secondary to an increase in PRL synthesis. Somatostatin (100 nM) inhibited the acute stimulatory effect of PRL-RF, but had no effect on the delayed stimulation of PRL release. On the other hand, cycloheximide (20 microM) completely inhibited the secondary increase in PRL release in response to PRL-RF but had no effect on the acute release. These results demonstrate that PRL-RF stimulates both the synthesis and release of decidual PRL.

Alloantigen presenting capacity of human decidual tissue

The capacity of first trimester human decidua-derived cells to serve as accessory cells for the presentation of alloantigens to unprimed T lymphocytes was assessed using a mixed lymphocyte culture (MLC) between accessory cell-depleted responder and stimulator peripheral blood lymphocytes (d PBL). In all of the seven experiments performed decidua-derived cells achieved significant reconstitution of the lymphoproliferative response between autologous responder and stimulator dPBL. Reconstitution indices (RIs) ranged between 3.1 and 22.2 and were significant in all cases. In six out of the seven experiments the level of lymphoproliferation exceeded that between untreated responder PBLs co-cultured with stimulator d PBL. Substitution of recombinant interleukin 1 (rIL-1) α or β for the decidua-derived cells resulted in significantly lower RIs than those achieved by the decidua-derived cells. These findings suggest that decidua-derived cells can serve as antigen presenting cells for the presentation of membrane-bound alloantigens in a primary lymphoproliferative response. This may have implications for maternal recognition of fetal antigens during first pregnancy.

Antigen presenting cells in human decidual tissue: III. Role of accessory cells in the activation of suppressor cells.

Human decidual antigen presenting cells (DAPCs) exposed to fetal cells in vitro induce generation of suppressor T cells among a population of peripheral blood lymphocytes. Human lymphocyte antigen (HLA)-class II positive antigen presenting cells were isolated from early normal human decidual tissue and from peripheral blood (PAPCs) by adhering Ficoll-Paque separated cell suspensions to fibronectin. In contrast to PAPCs, DAPCs pulsed with fetal antigens induced a radio-sensitive, Leu 1,2-positive T suppressor cell population. A nylon wool adherent B cell population is required during the in vitro induction of the suppressor cells. These suppressor cells impair primary mitogen and mixed lymphocyte culture (MLC) responses, generation of anti-trinitrophenyl (TNP) cytotoxic T lymphocytes, and antibody response of autologous and allogeneic lymphocytes. Only intact viable embryonic cells can effectively confer upon DAPCs the ability to induce T suppressor cells. The T suppressor cell induction by DAPCs primed with fetal antigens is restricted by the major histocompatibility complex. Our results show that the HLA-DR molecules are the most prominent restriction elements.

The effect of prostaglandin F 2α treatment on the endometrial oestradiol receptor system

The effects of prostaglandin F 2α treatment on oestradiol receptor levels were studied in human decidual tissues from the first trimester of pregnancy. Two types of nuclear oestradiol binding sites in decidual tissue were detected. Type I receptors bind the oestradiol with high affinity and low capacity (K d = 1.4 nM, B max = 0.3 pmol/mgDNA, Hill coeff = 1), while type II binding sites have lower affinity and higher capacity for the hormone (K d = 20.4 nM, B max = 2.0 pmol/mgDNA, Hill coeff = 3). The nuclear oestrogen binding capacities of decidual tissue were affected by prostaglandin F 2α in a time dependent manner. Within the first 10 hrs after prostaglandin F 2α application the concentration of low affinity, high capacity binding sites (type II) was enhanced in comparison with the control values. Later, 20–24 hrs after the treatment, both types of nuclear oestrogen binding sites were increased significantly. When prostaglandin F 2α treatment failed to induce abortion no change in the concentration of nuclear oestrogen binding sites was observed. Oestradiol receptor concentration in the cytoplasm apparently was not affected by prostaglandin F 2α treatment.

Progesterone and antiprogestins, a comparison of their effect on prostaglandin production by human secretory phase endometrium and decidua

The mechanism by which progesterone inhibits PG production is not clear. In systems using isolated human endometrial fragments, progesterone has been shown to inhibit PG production markedly. We have used such a system to test the action of two antiprogesterone steroids RU486 (Roussell-Uclaf) and ZK98734 (Schering) on isolated human endometrial and decidual tissue, with and without added progesterone. Progesterone (200nM) reduced PGF 2α production by the secretory phase endometrium from 10.9ng/mg tissue/24 hr to1.9 ng/mg/24hr on the third day of culture (p<01) and this effect was antagonised by the addition of either 200nM RU486 or 200nM ZK98734 (6.3 and 7.2 ng/mg /24 hr respectively). The antiprogestins on their own showed a slight inhibitory effect on day 3 and RU486 treatment resulted in a significant (p<.05) decrease in PG production from control. PGE and the main 13,14-dihydro-15-keto metabolites of E and F were also significantly decreased by progesterone and restored by the aMiprogestins. The PG production by decidua increased on days 2 and 3 in response to progesterone + antiprogestins but this was not significant. This data shows that the inhibition of PG production shown by progesterone, acting on secretory phase endometrium cultured as tissue fragments, is reversible by the receptor blocking antiprogestins.

Purification of decidual prolactin-releasing factor, a placental protein that stimulates prolactin release from human decidual tissue

Decidual prolactin-releasing factor (PRL-RF), a placental protein that stimulates the release of prolactin from human decidual tissue, has been purified from conditioned medium of human placental explants. The purification scheme consisted of ethanol extraction, anion exchange chromatography on DEAE-cellulose, size exclusion chromatography on Spherogel TSK-3000, and either a) immunoaffinity chromatography using an antiserum to a partially purified PRL-RF preparation or b) acetic acid-urea/SDS 2-dimensional PAGE. The apparent molecular weight of the purified releasing factor, estimated by SDS-PAGE, was 23,500 Da; and the half-maximal dose for the acute stimulation of prolactin release from human decidual cells was 0.05-0.1 ug/ml (2.2-4.4 nM).