107. INTERLEUKIN 11 AND LEUKEMIA INHIBITORY FACTOR REGULATE CYTOKINE NETWORKS IN HUMAN FIRST TRIMESTER PLACENTA

2009 ◽  
Vol 21 (9) ◽  
pp. 26
Author(s):  
E. Dimitriadis ◽  
P. Paiva ◽  
U. Manuelpillai ◽  
G. Weston ◽  
K. Meehan ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Leukemia Inhibitory Factor ◽  
First Trimester ◽  
Mechanisms Of Action ◽  
Cytokine Secretion ◽  
Inhibitory Factor ◽  
Trophoblast Invasion ◽  
Human Trophoblast ◽  
Vitro Model

Inadequate trophoblast invasion is thought to be involved in the pathogenesis of preeclampsia (PE). Shallow trophoblast invasion has been implicated to lead to placental hypoxia and a localised overproduction of pro-inflammatory cytokines. Interleukin (IL) 11 and leukemia inhibitory factor (LIF) are IL6-type cytokines produced at the maternal-fetal interphase and regulate human trophoblast migration/invasion, but their mechanisms of action are unknown. We aimed to determine the effect of hypoxia on human placental IL11/LIF secretion and the effect of IL11/LIF on placental cytokine secretion. As an in vitro model we cultured human first trimester placental villous explants (gestation weeks 7-9) in serum free conditions under either 2% (hypoxia) or 20% oxygen (normoxia) (N=8/gp) for 48h. Medium was assayed for IL11/LIF by ELISA. The effect of IL11/LIF (N=6/gp) on placental explant cytokine secretion (26 cytokines) was analysed using a quantitative multiplex immunoassay. Data was expressed as pg/mg wet weight and then as % change vs control (100%). IL11 secretion from explants was decreased by 76±5% (p<0.01) under hypoxia vs normoxia while LIF secretion did not differ significantly. Under normoxia the most highly abundant cytokines were IL6, IL8 and MCP-1 while moderate levels of G-CSF, CX3CL1, IP-10, and PGDF were present in conditioned medium. IL11 increased IL10 secretion while it decreased G-CSF, TNFa, IL1receptor antagonist (Ra), IL6 and PDGF secretion (p<0.05) vs control. Similarly LIF decreased G-CSF, TNFa, IL1Ra and additionally IL8 secretion (p<0.05) vs control. IL6 and VEGF secretion were increased (p<0.05) while MCP-1 was reduced (p<0.05) by hypoxia vs normoxia. This study identified for the first time that IL11 was reduced by hypoxia and identified a profile of cytokines secreted by placenta. IL11 and LIF regulated similar and unique anti-inflammatory cytokines in first trimester placenta. Whether placental IL11 or LIF is altered in women with PE remains to be elucidated however this study suggests mechanisms of action and confirms the importance of IL11/LIF in placentation.

scholarly journals Investigation of human trophoblast invasion in vitro

2020 ◽  
Vol 26 (4) ◽  
pp. 501-513 ◽  
Author(s):  
Yassen Abbas ◽  
Margherita Y Turco ◽  
Graham J Burton ◽  
Ashley Moffett
Keyword(s):  
Cell Lines ◽  
First Trimester ◽  
Trophoblast Cell ◽  
Extravillous Trophoblast ◽  
Trophoblast Invasion ◽  
Human Trophoblast ◽  
Uterine Environment ◽  
Microfluidic Assays ◽  
Invasion Assays

Abstract BACKGROUND In humans, inadequate trophoblast invasion into the decidua is associated with the ‘great obstetrical syndromes’ which include pre-eclampsia, foetal growth restriction (FGR) and stillbirth. The mechanisms regulating invasion remain poorly understood, although interactions with the uterine environment are clearly of central importance. Extravillous trophoblast (EVT) cells invade the uterus and transform the spiral arteries. Progress in understanding how they invade has been limited due to the lack of good in vitro models. Firstly, there are no non-malignant cell lines that have an EVT phenotype. Secondly, the invasion assays used are of limited use for the small numbers of primary EVT available from first-trimester placentas. We discuss recent progress in this field with the generation of new EVT lines and invasion assays using microfluidic technology. OBJECTIVE AND RATIONALE Our aim is to describe the established models used to study human trophoblast invasion in vivo and in vitro. The difficulties of obtaining primary cells and cell lines that recapitulate the phenotype of EVT are discussed together with the advantages and pitfalls of the different invasion assays. We compare these traditional end point assays to microfluidic assays where the dynamics of migration can be measured. SEARCH METHODS Relevant studies were identified by PubMed search, last updated on February 2020. A search was conducted to determine the number of journal articles published using the cell lines JEG-3, BeWo, JAR, HTR-8/Svneo, Swan-71 and primary human extravillous trophoblast in the last 5 years. OUTCOMES Deep trophoblast invasion into the maternal decidua is a particular feature of human pregnancy. This invasion needs to be finely regulated to allocate resources between mother and baby. A reliable source of EVT is needed to study in vitro how the uterine environment regulates this process. First, we critically discuss the issues with the trophoblast cell lines currently used; for example, most of them lack expression of the defining marker of EVT, HLA-G. Recently, advances in human stem cell and organoid technology have been applied to extraembryonic tissues to develop trophoblast cell lines that can grow in two (2D) and three dimensions (3D) and differentiate to EVT. This means that the ‘trophoblast’ cell lines currently in use should rapidly become obsolete. Second, we critically discuss the problems with assays to study trophoblast invasion. These lack physiological relevance and have simplified migration dynamics. Microfluidic assays are a powerful tool to study cell invasion because they require only a few cells, which are embedded in 3D in an extracellular matrix. Their major advantage is real-time monitoring of cell movement, enabling detailed analysis of the dynamics of trophoblast migration. WIDER IMPLICATIONS Trophoblast invasion in the first trimester of pregnancy remains poorly understood despite the importance of this process in the pathogenesis of pre-eclampsia, FGR, stillbirth and recurrent miscarriage. The new technologies described here will allow investigation into this critical process.

Endometrial leukemia inhibitory factor as a predictor of pregnancy after in vitro fertilization

2008 ◽  
Vol 102 (1) ◽  
pp. 23-27 ◽  
Author(s):  
Paulo Serafini ◽  
André M. Rocha ◽  
Cyntia T. Osório ◽  
Ismael da Silva ◽  
Eduardo L. Motta ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Leukemia Inhibitory Factor ◽  
Inhibitory Factor ◽  
Vitro Fertilization

Integrin switching regulates normal trophoblast invasion

Development ◽  
1994 ◽  
Vol 120 (12) ◽  
pp. 3657-3666 ◽  
Author(s):  
C.H. Damsky ◽  
C. Librach ◽  
K.H. Lim ◽  
M.L. Fitzgerald ◽  
M.T. McMaster ◽  
...  
Keyword(s):  
First Trimester ◽  
Uterine Wall ◽  
Trophoblast Invasion ◽  
Collagen Type Iv ◽  
Fibronectin Receptor ◽  
Type Iv ◽  
Invasive Capacity ◽  
Functional Consequences

Cells invade extracellular matrices in a regulated manner at specific times and places during normal development. A dramatic example is trophoblast invasion of the uterine wall. Previous studies have shown that differentiation of trophoblasts to an invasive phenotype is accompanied by temporally and spatially regulated switching of their integrin repertoire. In the first trimester human placenta, alpha 6 integrins are restricted to cytotrophoblast (CTB) stem cells and downregulated in invasive CTBs, whereas alpha 5 beta 1 and alpha 1 beta 1 integrins are upregulated in differentiating and invasive CTBs. The goal of the present study was to determine whether these changes have functional consequences for CTB invasiveness. Using an in vitro invasion model, we determined first that aggregates of invading first trimester CTBs in vitro undergo the same pattern of integrin switching as was observed in situ, thereby validating the utility of the model. We then showed that antibody perturbation of interactions involving laminin or collagen type IV and their integrin alpha 1/beta 1 receptor inhibited invasion by CTBs, whereas perturbing interactions between fibronectin and the alpha 5/beta 1 fibronectin receptor accelerated invasion. Finally, we report that later gestation CTBs, which display greatly decreased invasive capacity, are also unable to upregulate alpha 1 beta 1 complexes, providing further evidence that this integrin is critical for CTB invasion. This gestational regulation is transcriptional. These data indicate that integrin switching observed during differentiation in situ has significant functional consequences for CTB invasion. The data suggest further that differentiating CTBs upregulate counterbalancing invasion-accelerating and invasion-restraining adhesion mechanisms. We propose that this contributes to regulating the depth of CTB invasion during normal implantation.

Stem cell factor and leukemia inhibitory factor promote primordial germ cell survival by suppressing programmed cell death (apoptosis)

Development ◽  
1993 ◽  
Vol 118 (4) ◽  
pp. 1089-1094 ◽  
Author(s):  
M. Pesce ◽  
M.G. Farrace ◽  
M. Piacentini ◽  
S. Dolci ◽  
M. De Felici
Keyword(s):  
Cell Death ◽  
Stem Cell ◽  
Programmed Cell Death ◽  
Leukemia Inhibitory Factor ◽  
Stem Cell Factor ◽  
Tissue Transglutaminase ◽  
Primordial Germ Cell ◽  
Inhibitory Factor ◽  
High Level

Proliferating primordial germ cells (PGCs) isolated from mouse embryos soon after their arrival in the genital ridges would only survive in vitro at temperature of less than 30 degrees C (De Felici, M. and McLaren, A. (1983). Exp. Cell. Res. 144, 417–427; Wabik-Sliz, B. and McLaren, A. (1984). Exp. Cell. Res. 154, 530–536) or when co-cultured on cell feeder layers (Donovan, P. J., Stott, D., Godin, I., Heasman, J. and Wylie, C. C. (1986). Cell 44, 831–838; De Felici, M. and Dolci, S. (1991). Dev. Biol. 147, 281–284). In the present paper we report that mouse PGC death in vitro occurs with all the hallmarks of programmed cell death or apoptosis. We found that after 4–5 hours in culture many PGCs isolated from 12.5 dpc fetal gonads assumed a nuclear morphology and produced membrane bound fragments (apoptotic bodies) typical of apoptotic cells. In addition, PGCs in culture accumulated high level of tissue transglutaminase (tTGase; an enzyme that is induced and activated during apoptosis) and showed extensive degradation of DNA to oligonucleosomal fragments, which is characteristic of apoptosis. The physiological relevance of this mechanism of PGC death is supported by the finding that some PGCs undergoing apoptosis, as revealed by the high level of tTGase expression, were detected in the embryo. Most importantly, we show that the addition of stem cell factor (SCF) or leukemia inhibitory factor (LIF) to the culture medium, two cytokines known to favour PGC survival and/or proliferation in vitro, markedly reduced the occurrence of apoptosis in PGCs during the first hours in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Osteoblast and Osteoclast Activity Affect Bone Remodeling Upon Regulation by Mechanical Loading-Induced Leukemia Inhibitory Factor Expression in Osteocytes

2020 ◽  
Vol 7 ◽  
Author(s):  
Jingke Du ◽  
Jiancheng Yang ◽  
Zihao He ◽  
Junqi Cui ◽  
Yiqi Yang ◽  
...  
Keyword(s):  
Bone Remodeling ◽  
Mechanical Loading ◽  
Leukemia Inhibitory Factor ◽  
Fluid Shear Stress ◽  
Bone Structure ◽  
Inhibitory Factor ◽  
Tartrate Resistant Acid Phosphatase ◽  
Osteoclast Activity ◽  
Microgravity Simulation

PurposeBone remodeling is affected by mechanical stimulation. Osteocytes are the primary mechanical load-sensing cells in the bone, and can regulate osteoblast and osteoclast activity, thus playing a key role in bone remodeling. Further, bone mass during exercise is also regulated by Leukemia inhibitory factor (LIF). This study aimed to investigate the role of LIF in the mechanical response of the bone, in vivo and in vitro, and to elucidate the mechanism by which osteocytes secrete LIF to regulate osteoblasts and osteoclasts.MethodsA tail-suspension (TS) mouse model was used in this study to mimic muscular disuse. ELISA and immunohistochemistry were performed to detect bone and serum LIF levels. Micro-computed tomography (CT) of the mouse femurs was performed to measure three-dimensional bone structure parameters. Fluid shear stress (FSS) and microgravity simulation experiments were performed to study mechanical stress-induced LIF secretion and its resultant effects. Bone marrow macrophages (BMMs) and bone mesenchymal stem cells (BMSCs) were cultured to induce in vitro osteoclastogenesis and osteogenesis, respectively.ResultsMicro-CT results showed that TS mice exhibited deteriorated bone microstructure and lower serum LIF expression. LIF secretion by osteocytes was promoted by FSS and was repressed in a microgravity environment. Further experiments showed that LIF could elevate the tartrate-resistant acid phosphatase activity in BMM-derived osteoclasts through the STAT3 signaling pathway. LIF also enhanced alkaline phosphatase staining and osteogenesis-related gene expression during the osteogenic differentiation of BMSCs.ConclusionMechanical loading affected LIF expression levels in osteocytes, thereby altering the balance between osteoclastogenesis and osteogenesis.

scholarly journals Murine leukemia inhibitory factor enhances retroviral-vector infection efficiency of hematopoietic progenitors

Blood ◽  
1990 ◽  
Vol 76 (6) ◽  
pp. 1098-1103 ◽  
Author(s):  
FA Fletcher ◽  
DE Williams ◽  
C Maliszewski ◽  
D Anderson ◽  
M Rives ◽  
...  
Keyword(s):  
Leukemia Inhibitory Factor ◽  
Retroviral Vector ◽  
Inhibitory Factor ◽  
Hematopoietic Progenitors ◽  
Short Term ◽  
Infection Efficiency ◽  
Integration Sites ◽  
In Vitro Effects ◽  
Short Term Culture

Abstract We have investigated the in vitro effects of the cytokine leukemia inhibitory factor (LIF) on normal murine hematopoietic progenitors by measuring recovery and retroviral vector infection efficiency of 13-day posttransplant, spleen-colony-forming cell (CFU-S 13) in short-term culture. Up to a twofold increase in CFU-S13 recovery was observed, from 9.7 x 10(-5) cells in untreated controls to 17.8 to 19.5 x 10(-5) cells, depending on the concentration of LIF. Histologic analysis of spleen colonies from control and LIF-treated marrows demonstrated that there was no detectable alteration in the differentiative potential of CFU-S13. The efficiency of CFU-S13 infection was increased from 15% in untreated controls to 84% to 91% in LIF-treated marrows. Analysis of proviral integration sites in spleen colonies indicated that some CFU- S13 precursors were infected in the LIF-treated marrows.

scholarly journals Kidney-Replenishing Herb Induces SOCS-3 Expression via ERK/MAPK Pathway and Improves Growth of the First-Trimester Human Trophoblast Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-12
Author(s):  
Chang-ying Xing ◽  
De-xiang Zhang ◽  
Sui-qi Gui ◽  
Min-fang Tao
Keyword(s):  
Mapk Pathway ◽  
Recurrent Miscarriage ◽  
First Trimester ◽  
Mek Inhibitor ◽  
Biological Functions ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Mitogen Activated Protein ◽  
Human Trophoblast Cells

Kidney-replenishing herb is a traditional medicine formula in China which has been widely used for clinical treatment of recurrent miscarriage. Our previous study showed that Kidney-replenishing herb could promote proliferation and inhibit apoptosis of the human first-trimester trophoblasts. In the present study, we further explored the potential mechanism and signal pathway of Kidney-replenishing herb on human trophoblast cells. Our research showed that Kidney-replenishing herb stimulated proliferation and reduced apoptosis of human trophoblast cells in vitro, and this appeared to be positive correlation with SOCS-3 transcription, suggesting that Kidney-replenishing herb regulated biological functions of human trophoblast cells by inducing SCOS-3 expression. Furthermore, the Kidney-replenishing herb treatment stimulated the phosphorylation of ERK1/2, and blocking the signaling pathway by mitogen-activated protein MAPK (MEK) inhibitor, U0126, inhibited Kidney-replenishing herb-induced SOCS-3 transcription, depressed proliferation, and promoted apoptosis of human trophoblasts. Kidney-replenishing herbs still induced ERK1/2 phosphorylation after SOCS-3 siRNA silence. Overexpression of SOCS-3 stimulated the proliferation of trophoblast. These findings suggest that SOCS-3 expression is induced by Kidney-replenishing herbs via activation of MAPK pathways, and this may possibly be involved in promoting human trophoblast cells growth which is contributed to embryo development.

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