Publications by authors named "Udo R Markert"

114 Publications

Smoking for two- effects of tobacco consumption on placenta.

Mol Aspects Med 2021 Sep 11:101023. Epub 2021 Sep 11.

Placenta Lab, Department of Obstetrics, University Hospital Jena, Jena, Germany. Electronic address:

Tobacco smoking is an important public health issue recognized by the world health organization as one of the most serious, preventable risk factors for developing a series of pregnancy pathologies. Maternal smoking is positively associated with intrauterine growth restriction (IUGR) and gestational diabetes (GDM), but negatively associated with preeclampsia (PE). In this review, we examine epidemiological, clinical and laboratory studies of smoking effects on immunoregulation during pregnancy, trophoblast function, and placental vasculature development and metabolism. We aim to identify effects of tobacco smoke components on specific placental compartments or cells, which may contribute to the understanding of the influences of maternal smoking on placenta function in normal and pathological pregnancies. Data corroborates that in any trimester, smoking is unsafe for pregnancy and that its detrimental effects outweigh questionable benefits. The effects of maternal smoking on the maternal immune regulation throughout pregnancy and the impact of different tobacco products on fetal growth have not yet been fully understood. Smoking cessation rather than treatment with replacement therapies is recommended for future mothers because also single components of tobacco and its smoke may have detrimental effects on placental function.
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http://dx.doi.org/10.1016/j.mam.2021.101023DOI Listing
September 2021

Pregnancy and pandemics: Interaction of viral surface proteins and placenta cells.

Biochim Biophys Acta Mol Basis Dis 2021 11 24;1867(11):166218. Epub 2021 Jul 24.

Placenta Lab, Department of Obstetrics, Jena University Hospital, 07747 Jena, Germany.. Electronic address:

Throughout history, pandemics of infectious diseases caused by emerging viruses have spread worldwide. Evidence from previous outbreaks demonstrated that pregnant women are at high risk of contracting the diseases and suffering from adverse outcomes. However, while some viruses can cause major health complications for the mother and her fetus, others do not appear to affect pregnancy. Viral surface proteins bind to specific receptors on the cellular membrane of host cells and begin therewith the infection process. During pregnancy, the molecular features of these proteins may determine specific target cells in the placenta, which may explain the different outcomes. In this review, we display information on Variola, Influenza, Zika and Corona viruses focused on their surface proteins, effects on pregnancy, and possible target placental cells. This will contribute to understanding viral entry during pregnancy, as well as to develop strategies to decrease the incidence of obstetrical problems in current and future infections.
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http://dx.doi.org/10.1016/j.bbadis.2021.166218DOI Listing
November 2021

Inefficient Placental Virus Replication and Absence of Neonatal Cell-Specific Immunity Upon Sars-CoV-2 Infection During Pregnancy.

Front Immunol 2021 3;12:698578. Epub 2021 Jun 3.

Department of Obstetrics and Fetal Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.

Pregnant women have been carefully observed during the COVID-19 pandemic, as the pregnancy-specific immune adaptation is known to increase the risk for infections. Recent evidence indicates that even though most pregnant have a mild or asymptomatic course, a severe course of COVID-19 and a higher risk of progression to diseases have also been described, along with a heightened risk for pregnancy complications. Yet, vertical transmission of the virus is rare and the possibility of placental SARS-CoV-2 infection as a prerequisite for vertical transmission requires further studies. We here assessed the severity of COVID-19 and onset of neonatal infections in an observational study of women infected with SARS-CoV-2 during pregnancy. Our placental analyses showed a paucity of SARS-CoV-2 viral expression in term placentae under acute infection. No viral placental expression was detectable in convalescent pregnant women. Inoculation of placental explants generated from placentas of non-infected women at birth with SARS-CoV-2 revealed inefficient SARS-CoV-2 replication in different types of placental tissues, which provides a rationale for the low viral expression. We further detected specific SARS-CoV-2 T cell responses in pregnant women within a few days upon infection, which was undetectable in cord blood. Our present findings confirm that vertical transmission of SARS-CoV-2 is rare, likely due to the inefficient virus replication in placental tissues. Despite the predominantly benign course of infection in most mothers and negligible risk of vertical transmission, continuous vigilance on the consequences of COVID-19 during pregnancy is required, since the maternal immune activation in response to the SARS-CoV2 infection may have long-term consequences for children's health.
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http://dx.doi.org/10.3389/fimmu.2021.698578DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8211452PMC
June 2021

An obituary: Dr. Gérard Chaouat May 6, 1944 - April 23, 2021.

J Reprod Immunol 2021 May 13;145:103329. Epub 2021 May 13.

Department of Medical Biology, Medical School, University of Pecs, Pecs, Hungary. Electronic address:

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http://dx.doi.org/10.1016/j.jri.2021.103329DOI Listing
May 2021

Molecular characteristics of established trophoblast-derived cell lines.

Placenta 2021 05 4;108:122-133. Epub 2021 Mar 4.

Placenta Lab, Department of Obstetrics, Jena University Hospital, 07747, Jena, Germany. Electronic address:

Introduction: Research on human placental development and function lacks a conclusive in vivo model. To investigate the intracellular molecular mechanisms in trophoblast cells, different cell lines have been established during the last decades. So far, none of these accomplishes all features of primary trophoblast, thus their suitability as well as the transferability of the results has been discussed. The aim of this study is to assess molecular markers and features matching different trophoblast subpopulations in trophoblastic cell lines to provide orientation on their suitability and relevance for distinct research questions.

Methods: The commonly used trophoblastic cell lines, BeWo, JEG-3, HTR-8/SVneo, AC1-M59, AC1-M32, ACH-3P and Swan71 were selected. qPCR and immunoblotting were used to determine expression of characteristic molecular markers. C14MC, C19MC and miR-371-3 miRNA expression were investigated by real time PCR. Proliferation, migration and network stabilization assays were performed. Hormone secretion was determined by chemiluminescent-immunoassays. DNA profiles were obtained by Short Tandem Repeat (STR)-genotyping.

Results: Immortalized cell lines differ from choriocarcinoma-derived ones in the expression of HLA-G, E-cadherin, N-cadherin, VE-cadherin, cadherin-11, cytokeratin 7, vimentin, ADAM12 and PRG2. Compared to choriocarcinoma-derived cell lines, expression of C19MC and hormone secretion were almost absent in immortalized cell lines. Conversely, they express C14MC and exhibit higher migration and network stabilization.

Discussion: The data presented will help justify the use of a cell line to evaluate distinct features of trophoblast biology and pathology. In general, characteristics and markers of choriocarcinoma derived cell lines seem to be more similar to in vivo trophoblast than immortalized cell lines and thus might be regarded as more suitable models.
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http://dx.doi.org/10.1016/j.placenta.2021.02.022DOI Listing
May 2021

Influence of high glucose in the expression of miRNAs and IGF1R signaling pathway in human myometrial explants.

Arch Gynecol Obstet 2021 06 11;303(6):1513-1522. Epub 2021 Feb 11.

Laboratory of Reproductive and Extracellular Matrix Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.

Purpose: Several roles are attributed to the myometrium including sperm and embryo transport, menstrual discharge, control of uterine blood flow, and labor. Although being a target of diabetes complications, the influence of high glucose on this compartment has been poorly investigated. Both miRNAs and IGF1R are associated with diabetic complications in different tissues. Herein, we examined the effects of high glucose on the expression of miRNAs and IGF1R signaling pathway in the human myometrium.

Methods: Human myometrial explants were cultivated for 48 h under either high or low glucose conditions. Thereafter, the conditioned medium was collected for biochemical analyses and the myometrial samples were processed for histological examination as well as miRNA and mRNA expression profiling by qPCR.

Results: Myometrial structure and morphology were well preserved after 48 h of cultivation in both high and low glucose conditions. Levels of lactate, creatinine, LDH and estrogen in the supernatant were similar between groups. An explorative screening by qPCR arrays revealed that 6 out of 754 investigated miRNAs were differentially expressed in the high glucose group. Data validation by single qPCR assays confirmed diminished expression of miR-215-5p and miR-296-5p, and also revealed reduced miR-497-3p levels. Accordingly, mRNA levels of IGF1R and its downstream mediators FOXO3 and PDCD4, which are potentially targeted by miR-497-3p, were elevated under high glucose conditions. In contrast, mRNA expression of IGF1, PTEN, and GLUT1 was unchanged.

Conclusions: The human myometrium responds to short-term exposure (48 h) to high glucose concentrations by regulating the expression of miRNAs, IGF1R and its downstream targets.
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http://dx.doi.org/10.1007/s00404-020-05940-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8087607PMC
June 2021

Research on nanoparticles in human perfused placenta: State of the art and perspectives.

Placenta 2021 01 28;104:199-207. Epub 2020 Dec 28.

Laboratory for Particles-Biology Interactions, Empa, Swiss Federal Laboratories for Materials Science and Technology, Lerchenfeldstrasse 5, 9014, St. Gallen, Switzerland. Electronic address:

Increasing human exposure to nanoparticles (NPs) from various sources raises concerns for public health, especially for vulnerable risk groups like pregnant women and their developing fetuses. However, nanomedicine and the prospect of creating safe and effective NP-based formulations of drugs hold great promise to revolutionize treatment during pregnancy. With maternal and fetal health at stake, risks and opportunities of NPs in pregnancy need to be carefully investigated. Importantly, a comprehensive understanding of NP transport and effects at the placenta is urgently needed considering the central position of the placenta at the maternal-fetal interface and its many essential functions to enable successful pregnancy. The perfusion of human placental tissue provides a great opportunity to achieve predictive human relevant insights, circumventing uncertainties due to considerable differences in placental structure and function across species. Here, we have reviewed the current literature on the ex vivo human placenta perfusion of NPs. From 16 available studies, it was evident that placental uptake and transfer of NPs are highly dependent on their characteristics like size and surface modifications, which is in line with previous observations from in vitro and animal transport studies. These studies further revealed that special considerations apply for the perfusion of NPs and we identified relevant controls that should be implemented in future perfusion studies. While current studies mostly focused on placental transfer of NPs to conclude on potential fetal exposure, the ex vivo placental perfusion model has considerable potential to reveal novel insights on NP effects on placental tissue functionality and signaling that could indirectly affect maternal-fetal health.
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http://dx.doi.org/10.1016/j.placenta.2020.12.014DOI Listing
January 2021

Role of IL-36 Cytokines in the Regulation of Angiogenesis Potential of Trophoblast Cells.

Int J Mol Sci 2020 Dec 30;22(1). Epub 2020 Dec 30.

Placenta Lab, Department of Obstetrics, University Hospital Jena, 07740 Jena, Germany.

IL-36 cytokines (the agonists IL-36α, IL-36β, IL-36γ, and the antagonist IL-36Ra) are expressed in the mouse uterus and associated with maternal immune response during pregnancy. Here, we characterize the expression of IL-36 members in human primary trophoblast cells (PTC) and trophoblastic cell lines (HTR-8/SVneo and JEG-3) and upon treatment with bacterial and viral components. Effects of recombinant IL-36 on the migration capacity of trophoblastic cells, their ability to interact with endothelial cells and the induction of angiogenic factors and miRNAs (angiomiRNAs) were examined. Constitutive protein expression of IL-36 (α, β, and γ) and their receptor (IL-36R) was found in all cell types. In PTC, transcripts for all IL-36 subtypes were found, whereas in trophoblastic cell lines only for and . A synthetic analog of double-stranded RNA (poly I:C) and lipopolysaccharide (LPS) induced the expression of IL-36 members in a cell-specific and time-dependent manner. In HTR-8/SVneo cells, IL-36 cytokines increased cell migration and their capacity to interact with endothelial cells. and mRNA and protein, as well as the angiomiRNAs miR-146a-3p and miR-141-5p were upregulated as IL-36 response in PTC and HTR-8/SVneo cells. In conclusion, IL-36 cytokines are modulated by microbial components and regulate trophoblast migration and interaction with endothelial cells. Therefore, a fundamental role of these cytokines in the placentation process and in response to infections may be expected.
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http://dx.doi.org/10.3390/ijms22010285DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7794747PMC
December 2020

Introduction to the special issue on extracellular vesicles and reproduction.

Am J Reprod Immunol 2021 02 26;85(2):e13387. Epub 2021 Jan 26.

Department of Pediatrics, Women and Infants Hospital, Warren Alpert Medical School of Brown University, Providence, RI, USA.

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http://dx.doi.org/10.1111/aji.13387DOI Listing
February 2021

Immunomodulatory properties of extracellular vesicles in the dialogue between placental and immune cells.

Am J Reprod Immunol 2021 02 27;85(2):e13383. Epub 2020 Dec 27.

Placenta Lab, Department of Obstetrics, Jena University Hospital, Jena, Germany.

Extracellular vesicle (EV)-mediated communication has been implicated in the cooperative alliance between trophoblast and immune cells toward maternal tolerance and placentation. Syncytiotrophoblast cells secrete EVs directly into the maternal circulation, which are taken up by immune cells, endothelial cells, and other cell types. Initial evidence also shows that EVs produced by immune cells are, in turn, incorporated by trophoblast cells and modulate placental responses. Non-coding RNAs (ncRNAs), proteins, and lipid mediators transported in EVs are able to influence proliferation, differentiation, cytokine production, and immunological responses of recipient cells. The molecular alphabet and cellular targets involved in this dialogue are being revealed. Nevertheless, several questions regarding the whole content, surface markers, and biological functions of EVs still remain to be investigated in both physiological and pathological conditions. Analysis of circulating EVs in maternal blood has the potential to serve as a minimally invasive approach to monitoring placental functions and immunological features of pregnancy, aiding in the diagnostics of complications. This review addresses the immunomodulatory properties of EVs and their tasks in the communication between placental and immune cells.
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http://dx.doi.org/10.1111/aji.13383DOI Listing
February 2021

Placental miRNAs in feto-maternal communication mediated by extracellular vesicles.

Placenta 2020 12 11;102:27-33. Epub 2020 Jul 11.

Placenta Lab, Department of Obstetrics, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany. Electronic address:

A complex network composed of at least 1900 microRNA (miRNA) species orchestrates the development and function of the human placenta. These molecules regulate genes and pathways operating major functional processes in trophoblast cells such as proliferation, invasion, differentiation, and metabolism. Nevertheless, the cellular localization and role of most placental miRNAs remain to be determined. The existence of eutherian- (C14MC) and primate-specific miRNA clusters (C19MC), together with human placenta-specific miRNAs, indicate the relevance of these molecules in evolution and diversification of the placenta, including the acquisition of its unique features in humans. They may be related also to diseases that are exclusively present in primates, such as preeclampsia. Changes in the miRNA expression profile have been reported in several placental pathologies. Which miRNAs are involved in the pathomechanism of these diseases or act to maintain placental homeostasis is uncertain. Placenta-derived miRNAs are packed into extracellular vesicles (EVs) and distributed through the maternal circulation to distant organs, where they contribute to adaptations required during pregnancy. Similarly, the placenta also receives molecular information from other tissues to adapt fetoplacental metabolic demands to the maternal energetic supply. These processes can be impaired in pathologic conditions. Therefore, the collection of circulating placental miRNAs constitutes potentially a minimally-invasive approach to assess the fetoplacental status and to diagnose pregnancy diseases. Future therapies may include manipulation of miRNA levels for prevention and treatment of placental complications to protect maternal health and fetal development.
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http://dx.doi.org/10.1016/j.placenta.2020.07.001DOI Listing
December 2020

Enrichment and characterization of extracellular vesicles from ex vivo one-sided human placenta perfusion.

Am J Reprod Immunol 2021 08 28;86(2):e13377. Epub 2020 Nov 28.

Placenta-Lab, Department of Obstetrics, Jena University Hospital, Jena, Germany.

Problem: Extracellular vesicles (EVs) released by the placenta are packed with biological information and play a major role in fetomaternal communication. Here, we describe a comprehensive set-up for the enrichment and characterization of EVs from human placenta perfusion and their application in further assays.

Method Of Study: Human term placentas were used for 3 h ex vivo one-sided perfusions to simulate the intervillous circulation. Thereafter, populations of small (sEVs) and large EV (lEVs) were enriched from placental perfusate via serial ultracentrifugation. Following, EV populations were characterized regarding their size, protein concentration, RNA levels, expression of surface markers as well as their uptake and miRNA transfer to recipient cells.

Results: The sEV and lEV fractions from an entire perfusate yielded, respectively, 294 ± 32 µg and 525 ± 96 µg of protein equivalents and 2.6 ± 0.5 µg and 3.6 ± 0.9 µg of RNA. The sEV fraction had a mean diameter of 117 ± 47 nm, and the lEV fraction presented 236 ± 54 nm. CD63 was strongly detected by dot blot in sEVs, whereas only traces of this marker were found in lEVs. Both EV fractions were positive for the trophoblast marker PLAP (placental alkaline phosphatase) and annexin A1. EV internalization in immune cells was visualized by confocal microscopy, and the transfer of placental miRNAs was detected by quantitative real-time PCR (qPCR).

Conclusions: Enriched EV populations showed characteristic features of sEVs and lEVs. EV uptake and transfer of miRNAs to recipient cells demonstrated their functional integrity. Therefore, we advocate the ex vivo one-sided placenta perfusion as a robust approach for the collection of placental EVs.
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http://dx.doi.org/10.1111/aji.13377DOI Listing
August 2021

Cytogenomics of six human trophoblastic cell lines.

Placenta 2021 01 13;103:72-75. Epub 2020 Oct 13.

Jena University Hospital, Placenta-Lab, Department of Obstetrics, Am Klinikum 1, D-07747, Jena, Germany.

Trophoblastic cell lines are established models used to examine human placenta physiology and disease. We performed concurrent cytogenetic analyses of six established and well-studied trophoblastic cell lines including JAR, BeWo, JEG-3, AC-1M59, HTR8/SVneo, and ACH-3P. All cell lines showed near triploid or tetraploid karyotypes with unique inter- and intra-clonal aberrations, which result possibly from long-term culture or defects in the placenta or its malignant choriocarcinoma origin. Variable aneuploidy in 'standard' cell lines is under-appreciated and may not reflect the in vivo situation. It has the potential to negatively impact our understanding of normal cell function and cause disagreement between studies.
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http://dx.doi.org/10.1016/j.placenta.2020.10.011DOI Listing
January 2021

Inspired by the human placenta: a novel 3D bioprinted membrane system to create barrier models.

Sci Rep 2020 09 24;10(1):15606. Epub 2020 Sep 24.

Cellbricks GmbH, Berlin, 13355, Germany.

Barrier organ models need a scaffold structure to create a two compartment culture. Technical filter membranes used most often as scaffolds may impact cell behaviour and present a barrier themselves, ultimately limiting transferability of test results. In this work we present an alternative for technical filter membrane systems: a 3D bioprinted biological membrane in 24 well format. The biological membrane, based on extracellular matrix (ECM), is highly permeable and presents a natural 3D environment for cell culture. Inspired by the human placenta we established a coculture of a trophoblast-derived cell line (BeWo b30), together with primary placental fibroblasts within the biological membrane (simulating villous stroma) and primary human placental endothelial cells-representing three cellular components of the human placental villus. All cell types maintained their cell type specific marker expression after two weeks of coculture on the biological membrane. In permeability assays the trophoblast layer developed a barrier on the biological membrane, which was even more pronounced when cocultured with fibroblasts. In this work we present a filter membrane free scaffold, we characterize its properties and assess its suitability for cell culture and barrier models. Further we show a novel placenta inspired model in a complex bioprinted coculture. In the absence of an artificial filter membrane, we demonstrate barrier architecture and functionality.
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http://dx.doi.org/10.1038/s41598-020-72559-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7515925PMC
September 2020

Beyond Uterine Natural Killer Cell Numbers in Unexplained Recurrent Pregnancy Loss: Combined Analysis of CD45, CD56, CD16, CD57, and CD138.

Diagnostics (Basel) 2020 Aug 29;10(9). Epub 2020 Aug 29.

Placenta Lab, Department of Obstetrics, Jena University Hospital, 07747 Jena, Germany.

Changes in the number and cytotoxic potential of uterine Natural Killer (uNK) cells have been associated with reduced fertility. To provide a better characterization of immunophenotypes in the endometrium of women with uRPL (unexplained recurrent pregnancy loss), we examined the applicability of a set of five immune cell markers. The concentration (cells/mm) of CD45 leukocytes, CD56 uNK cells, and CD138 plasma cells as well as of CD16 and CD57 cells, which indicate high cytotoxic uNK cells, were assessed by immunohistochemistry in endometrial biopsies from 61 uRPL patients and 10 controls. Control fertile endometria presented 90-300 CD56 uNK cells/mm. uRPL cases were classified in subgroups of low (uRPL-CD56 < 90 cells/mm), normal (uRPL-CD56 90-300 cells/mm), and high uNK cell counts (uRPL-CD56 > 300 cells/mm). Some cases from the uRPL-CD56 and uRPL-CD56 subgroups showed elevated proportions of cytotoxic CD16 and CD57 cells in relation to CD56 cells. In the uRPL-CD56 subgroup, the CD57/CD56 ratio was reduced in most samples and the CD16/CD56 ratio was unaltered. Analysis of CD138 excluded the influence of chronic endometritis on these observations. Our results reinforce a link between uRPL and a dysfunctional endometrial environment associated with distinct immune cell profiles.
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http://dx.doi.org/10.3390/diagnostics10090650DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555015PMC
August 2020

Are uterine natural killer and plasma cells in infertility patients associated with endometriosis, repeated implantation failure, or recurrent pregnancy loss?

Arch Gynecol Obstet 2020 12 14;302(6):1487-1494. Epub 2020 Jul 14.

Department of Obstetrics, Gynecology and REI (UniKiD), Medical Faculty, Medical Center University of Düsseldorf, University Hospital Düsseldorf, Moorenstr. 5, 40225, Düsseldorf, Germany.

Purpose: Infertility is a debilitating situation that millions of women around the world suffer from, but the causal relationship between infertility and endometriosis is still unclear. We hypothesize that the immune cell populations of uterine natural killer cells (uNK) and plasma cells (PC) which define chronic endometritis could differ in patients with or without endometriosis and therefore be the link to endometriosis-associated infertility.

Methods: Our retrospective study includes 173 patients that underwent an endometrial scratching in the secretory phase of the menstrual cycle and subsequently immunohistochemical examination for uNK cells and PC. Sixty-seven patients were diagnosed with endometriosis, 106 served as the control cohort.

Results: The risk for an elevated number of uNK cells in women with endometriosis is not increased as compared to the control group. Our findings suggest that patients with endometriosis are 1.3 times more likely to have chronic endometritis (CE) as compared to those without and that the treatment with doxycycline might increase pregnancy rates. Endometriosis and an increased number of uNK cells seem to be unrelated.

Conclusions: In contrast to the lately published connection between endometriosis, infertility and increased uNK cells, we could not find any evidence that patients with endometriosis are more prone to elevated uterine uNK cells. Counting of PC in endometrial biopsies might be a new approach in the search of biomarkers for the nonsurgical diagnosis of endometriosis since our findings suggest a connection.
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http://dx.doi.org/10.1007/s00404-020-05679-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584523PMC
December 2020

Doxorubicin induces cytotoxicity and miR-132 expression in granulosa cells.

Reprod Toxicol 2020 Jun 4;96:95-101. Epub 2020 Jun 4.

Placenta Lab, Department of Obstetrics, Jena University Hospital, Jena, Germany.

Doxorubicin (DOX) is one of the most commonly used drugs for the treatment of childhood cancers, including leukemia and lymphomas. Despite the high survival rate, female leukemia survivors are at higher risk of ovarian failure and infertility later in life. Treatment with chemotherapeutic drugs like DOX is associated with damage in ovarian follicles, but the affectation grade of granulosa cells remains unclear. To assess and avoid the possible side-effects of DOX, early biomarkers of ovarian injury and chemotherapy-induced ovarian toxicity should be identified. MicroRNAs (miRNAs) have emerged in recent years as a promising new class of biomarkers for drug-induced tissue toxicity. In this study, the effects of DOX on cell viability, steroidogenesis, and miRNA expression were studied in primary granulosa cells (GCs) and in two cellular models (COV434 and KGN cells). We report that compared to other chemotherapeutic drugs, DOX treatment is more detrimental to granulosa cells as observed by decrease of cell viability. Treatment with DOX changes the expression of the aromatase gene (CYP19A1) and the secretion of 17β-estradiol (E2) in a cell-specific manner. miR-132-3p is dose-dependently increased by DOX in all cellular models. In absence of DOX, miR-132-3p overexpression in COV434 cells has no effect on E2 secretion or CYP19A1 expression. Altogether, these findings contribute to understanding the hormonal disbalance caused by DOX in human ovarian cells and suggest miR-132 as a putative sensor to predict DOX-induced ovarian toxicity.
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http://dx.doi.org/10.1016/j.reprotox.2020.06.001DOI Listing
June 2020

MiR-519d-3p in Trophoblastic Cells: Effects, Targets and Transfer to Allogeneic Immune Cells via Extracellular Vesicles.

Int J Mol Sci 2020 May 14;21(10). Epub 2020 May 14.

Placenta Lab, Department of Obstetrics, Jena University Hospital, 07740 Jena, Germany.

Members of the placenta-specific miRNA cluster C19MC, including miR-519d, are secreted by fetal trophoblast cells within extracellular vesicles (EVs). Trophoblast-derived EVs can be internalized by the autologous trophoblast and surrounding maternal immune cells, resulting in coordination of cellular responses. The study of functions and targets of placental miRNAs in the donor and recipient cells may contribute to the understanding of the immune tolerance essential in pregnancy. Here, we report that miR-519d-3p levels correlate positively with cell proliferation and negatively with migration in trophoblastic cell lines. Inhibition of miR-519d-3p in JEG-3 cells increases caspase-3 activation and apoptosis. PDCD4 and PTEN are targeted by miR-519d-3p in a cell type-specific manner. Transfection of trophoblastic cell lines with miR-519d mimic results in secretion of EVs containing elevated levels of this miRNA (EV). Autologous cells enhance their proliferation and decrease their migration ability when treated with EV. NK92 cells incorporate EV-delivered miR-519d-3p at higher levels than Jurkat T cells. EV increases the proliferation of Jurkat T cells but decreases that of NK92 cells. Altogether, miR-519d-3p regulates pivotal trophoblast cell functions, can be transferred horizontally via EVs to maternal immune cells and exerts functions therein. Vesicular miRNA transfer from fetal trophoblasts to maternal immune cells may contribute to the immune tolerance in pregnancy.
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http://dx.doi.org/10.3390/ijms21103458DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7278925PMC
May 2020

Evaluation of peripheral and uterine immune status of chronic endometritis in patients with recurrent reproductive failure.

Fertil Steril 2020 01 9;113(1):187-196.e1. Epub 2019 Nov 9.

Shenzhen Key Laboratory of Reproductive Immunology for Periimplantation, Shenzhen Zhongshan Institute for Reproduction and Genetics, Shenzhen Zhongshan Urology Hospital, Shenzhen, Guangdong, People's Republic of China. Electronic address:

Objective: To investigate whether chronic endometritis (CE) affects the immune status of peripheral blood and endometrium in patients with recurrent reproductive failure (RRF).

Design: Retrospective study.

Setting: Private fertility center.

Patients(s): A total of 524 RRF patients, including 324 women with recurrent miscarriage (RM) and 200 women with recurrent implantation failure (RIF).

Intervention(s): Peripheral blood and endometrium samples were collected in the midluteal phase before in vitro fertilization treatment or pregnancy. The number of peripheral T, natural killer (NK), and B cells, as well as cytotoxicity of NK cells and expression of T1 cytokines were analyzed with the use of flow cytometry, and uterine immune cells were subjected to immunohistochemistry.

Main Outcome Measure(s): Peripheral immune cells, cytokines, NK cytotoxicity, and endometrial immune cells were compared in RRF patients with versus without CE.

Result(s): The proportion and function of the analyzed immune cell subsets in peripheral blood as well as the percentages of CD56 NK cells, CD163 M2 macrophages, and CD1a immature dendritic cells in the endometrium were not significantly altered between non-CE and CE patients, whereas the proportions of uterine CD68 macrophages, CD83 mature dendritic cells, CD8 T cells, and Foxp3 regulatory T cells were significantly elevated in CE patients. After antibiotic treatment, the percentage of CD68 macrophages, CD83 mature dendritic cells, CD8 T cells, and Foxp3 regulatory T cells in endometrium were significantly reduced in patients with cured CE.

Conclusion(s): CE contributes to elevated endometrial infiltration levels of immune cells. The excessive presence of endometrial immune cells in CE patients may be involved in reduced endometrial receptivity and recurrent pregnancy failures.
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http://dx.doi.org/10.1016/j.fertnstert.2019.09.001DOI Listing
January 2020

Identification of miRNAs and associated pathways regulated by Leukemia Inhibitory Factor in trophoblastic cell lines.

Placenta 2019 12 12;88:20-27. Epub 2019 Sep 12.

Placenta-Labor, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany. Electronic address:

Introduction: Leukemia Inhibitory Factor (LIF) regulates behavior of trophoblast cells and their interaction with immune and endothelial cells. In vitro, trophoblast cell response to LIF may vary depending on the cell model. Reported differences in the miRNA profile of trophoblastic cells may be responsible for these observations. Therefore, miRNA expression was investigated in four trophoblastic cell lines under LIF stimulation followed by in silico analysis of altered miRNAs and their associated pathways.

Methods: Low density TaqMan miRNA assays were used to quantify levels of 762 mature miRNAs under LIF stimulation in three choriocarcinoma-derived (JEG-3, ACH-3P and AC1-M59) and a trophoblast immortalized (HTR-8/SVneo) cell lines. Expression of selected miRNAs was confirmed in primary trophoblast cells and cell lines by qPCR. Targets and associated pathways of the differentially expressed miRNAs were inferred from the miRTarBase followed by a KEGG Pathway Enrichment Analysis. HTR-8/SVneo and JEG-3 cells were transfected with miR-21-mimics and expression of miR-21 targets was assessed by qPCR.

Results: A similar number of miRNAs changed in each tested cell line upon LIF stimulation, however, low coincidence of individual miRNA species was observed and occurred more often among choriocarcinoma-derived cells (complete data set at http://www.ncbi.nlm.nih.gov/geo/ under GEO accession number GSE130489). Altered miRNAs were categorized into pathways involved in human diseases, cellular processes and signal transduction. Six cascades were identified as significantly enriched, including JAK/STAT and TGFB-SMAD. Upregulation of miR-21-3p was validated in all cell lines and primary cells and STAT3 was confirmed as its target.

Discussion: Dissimilar miRNA responses may be involved in differences of LIF effects on trophoblastic cell lines.
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http://dx.doi.org/10.1016/j.placenta.2019.09.005DOI Listing
December 2019

Trastuzumab in the Treatment of Pregnant Breast Cancer Patients - an Overview of the Literature.

Geburtshilfe Frauenheilkd 2019 Jun 14;79(6):618-625. Epub 2019 Jun 14.

Universitätsklinikum Jena, Klinik für Geburtsmedizin, Placenta-Labor, Jena, Germany.

Breast cancer is one of the most common malignancies which appear during pregnancy. Since women are increasingly not giving birth until they are at a more advanced age, it can be assumed that the incidence of pregnancy-related breast cancers will continue to increase in the future. Because of pregnancy-induced changes and conservative diagnosis, these carcinomas are frequently not detected until they are at an advanced stage and thus generally require systemic adjuvant therapy. The available data on optimal chemotherapeutic management are limited. Particularly for the use of the target agent trastuzumab which could crucially contribute to improving the prognosis in the therapy of HER2-overexpressing breast cancer in non-pregnant women, there is a lack of definitive information regarding the profile of action and safety in pregnancy as well as with regard to any long-term effects on the child. Thirty-eight pregnancies on trastuzumab for the treatment of breast cancer were able to be analysed in the literature currently available. Information can be gained from this and conclusions can be drawn which can individualise and decisively improve therapeutic options in the future for the pregnant breast cancer patient.
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http://dx.doi.org/10.1055/a-0880-9295DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570610PMC
June 2019

IL-36 Cytokines: Regulators of Inflammatory Responses and Their Emerging Role in Immunology of Reproduction.

Int J Mol Sci 2019 Apr 3;20(7). Epub 2019 Apr 3.

Placenta Lab, Department of Obstetrics, Jena University Hospital, 07740 Jena, Germany.

The IL-36 subfamily of cytokines has been recently described as part of the IL-1 superfamily. It comprises three pro-inflammatory agonists (IL-36α, IL-36β, and IL-36γ), their receptor (IL-36R), and one antagonist (IL-36Ra). Although expressed in a variety of cells, the biological relevance of IL-36 cytokines is most evident in the communication between epithelial cells, dendritic cells, and neutrophils, which constitute the common triad responsible for the initiation, maintenance, and expansion of inflammation. The immunological role of IL-36 cytokines was initially described in studies of psoriasis, but novel evidence demonstrates their involvement in further immune and inflammatory processes in physiological and pathological situations. Preliminary studies have reported a dynamic expression of IL-36 cytokines in the female reproductive tract throughout the menstrual cycle, as well as their association with the production of immune mediators and cellular recruitment in the vaginal microenvironment contributing to host defense. In pregnancy, alteration of the placental IL-36 axis has been reported upon infection and pre-eclampsia suggesting its pivotal role in the regulation of maternal immune responses. In this review, we summarize current knowledge regarding the regulatory mechanisms and biological actions of IL-36 cytokines, their participation in different inflammatory conditions, and the emerging data on their potential role in normal and complicated pregnancies.
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http://dx.doi.org/10.3390/ijms20071649DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6479377PMC
April 2019

The immunology of the macaque placenta: A detailed analysis and critical comparison with the human placenta.

Crit Rev Clin Lab Sci 2019 03 11;56(2):118-145. Epub 2019 Jan 11.

b Placenta Lab, Department of Obstetrics , University Hospital Jena , Jena , Germany.

The cynomolgus monkey is increasingly considered in toxicological research as the most appropriate model for humans due to the species' close physiological contiguity, including reproductive physiology. Here, literature on the cynomolgus monkey placenta is reviewed in regards to its similarity to the human placenta and particularly for its immunological role, which is not entirely mirrored in humans. Pertinent original data are included in this article. The cynomolgus monkey placenta is evaluated based on three aspects: first, morphological development; second, the spatial and temporal appearance of maternal and fetal immune cells and certain immune cell products of the innate and adaptive immune systems; and third, the expression of relevant immune tolerance-related molecules including the homologs of anti-human leucocyte antigen, indoleamine 2,3-dioxygenase, FAS/FAS-L, annexin II, and progesterone. Parameters relevant to the immunological role of the placenta are evaluated from the immunologically immature stage of gestational day (GD) 50 until more mature stages close to birth. Selected comparisons are drawn with human and other laboratory animal placentas. In conclusion, the cynomolgus monkey placenta has a high degree of morphological and physiological similarity to the human placenta. However, there are differences in the topographical distribution of cell types and immune tolerance-related molecules. Three basic features are recognized: (1) the immunological capacity of the placenta changes throughout the lifetime of the organ; (2) these immunological changes include multiple parameters such as morphological adaptations, cell type involvement, and changes in immune-relevant molecule expression; and (3) the immune systems of two genetically disparate individuals (mother and child) are functionally intertwined at the maternal-fetal interface.
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http://dx.doi.org/10.1080/10408363.2018.1538200DOI Listing
March 2019

Zika virus infection in human placental tissue explants is enhanced in the presence of dengue virus antibodies in-vitro.

Emerg Microbes Infect 2018 Dec 1;7(1):198. Epub 2018 Dec 1.

Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.

The current Zika virus (ZIKV) outbreak is associated with neurological malformations and disorders in neonates. Areas of increased incidence of malformations may overlap with dengue-hyperendemic areas. ZIKV infection is enhanced by antibodies against dengue virus (DENV) in cell culture and inbred mice. Sufficiently powered clinical studies or primate studies addressing the enhancement of fetal ZIKV infection after previous dengue infection are not available. The human placenta is susceptible to ZIKV in vitro, but it is unknown whether antibody-dependent enhancement of ZIKV infection occurs at the placental barrier. Here we studied ZIKV infection in placental tissue in the presence of DENV-immune sera. Explants from the amniochorionic membrane, the chorionic villi, and the maternal decidua were infected with ZIKV in the presence of DENV type 1-, 2-, or 4-immune sera, or controls. Presence of DENV antibodies of any type enhanced the percentage of successful infections of organ explants between 1.42- and 2.67-fold, and led to a faster replication as well as significantly increased virus production. No enhancement was seen with yellow fever or chikungunya virus control sera. Pre-existing DENV antibodies may pose an increased risk of trans-placental ZIKV transmission.
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http://dx.doi.org/10.1038/s41426-018-0199-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6274641PMC
December 2018

Correction: Placenta - Worth Trying? Human Maternal Placentophagy: Possible Benefit and Potential Risks.

Geburtshilfe Frauenheilkd 2018 09 16;78(9):e1. Epub 2018 Oct 16.

Plazenta-Labor, Klinik für Geburtsmedizin, Universitätsklinikum Jena, Jena, Germany.

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http://dx.doi.org/10.1055/a-0753-5753DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6191299PMC
September 2018

Placenta - Worth Trying? Human Maternal Placentophagia: Possible Benefit and Potential Risks.

Geburtshilfe Frauenheilkd 2018 Sep 14;78(9):846-852. Epub 2018 Sep 14.

Plazenta-Labor, Klinik für Geburtsmedizin, Universitätsklinikum Jena, Jena, Germany.

The use of placenta preparations as an individual puerperal remedy can be traced back to historical, traditional practices in Western and Asian medicine. To evaluate the ingestion of processed placenta as a puerperal remedy, the potential risks (trace elements, microorganisms) and possible benefit (hormones in the placental tissue) of such a practice are discussed in this article based on a literature review.
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http://dx.doi.org/10.1055/a-0674-6275DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6138470PMC
September 2018

Comparison of dienogest effects upon 3,3'-diindolylmethane supplementation in models of endometriosis and clinical cases.

Reprod Biol 2018 Sep 9;18(3):252-258. Epub 2018 Jul 9.

NAVAD LIFE SCIENCES PTE. LTD, Singapore. Electronic address:

Dienogest (DNG) administration is a well-established treatment for endometriosis but bleeding irregularities remain its main disadvantage. Changes in diet, mainly to vegetable consumption, are beneficial in the treatment of estrogen-related pathologies but their use for endometriosis has been poorly studied. In this study, addition of the phytochemical 3,3'-diindolylmethane (DIM) to DNG therapy has been investigated in in vitro and ex vivo models for endometriosis and in a small cohort of women with endometriosis. Endometrial Ishikawa cells were treated with DNG or DIM at dosages from 10 M to 10 M for up to 72 h. Cell proliferation was measured by assessing BrdU incorporation. Endometrial tissue from women with endometriosis and controls was incubated with DNG or a combination of DNG and DIM. Tissue viability was determined using a modified colorimetric MTS assay. 17β-estradiol secretion was quantified by an electro-chemiluminescence immunoassay. Finally, DNG as monotherapy or in combination with DIM was randomly administered to women with endometriosis (n = 8) over 3 months. Bleeding patterns and associated pelvic pain were assessed by Visual Analogue Scale (VAS). DNG and DIM significantly reduced cell proliferation in Ishikawa cells. Ex vivo, DIM reduced viability and estradiol secretion specifically in endometriotic but not in normal endometrial tissue. This effect was enhanced by combination with DNG. Endometriosis associated pelvic pain was significantly reduced in patients taking the DNG-DIM combination therapy compared to those taking DNG alone. Bleeding pattern (number and duration of episodes) was significantly improved by addition of DIM to the DNG treatment. In conclusion, addition of DIM enhances effects of DNG ex vivo and may ameliorate bleeding patterns in endometriosis patients.
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http://dx.doi.org/10.1016/j.repbio.2018.07.002DOI Listing
September 2018

Human placentophagy: Effects of dehydration and steaming on hormones, metals and bacteria in placental tissue.

Placenta 2018 07 16;67:8-14. Epub 2018 May 16.

Placenta-Lab, Department of Obstetrics, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany. Electronic address:

Introduction: Human maternal placentophagy, the behavior of ingesting the own raw or processed placenta postpartum, is a growing trend by women of western societies. This study aims to identify the impact of dehydration and steaming on hormone and trace element concentration as well as microbial contamination of placental tissue.

Methods: A total of nine placentas have been processed: six were studied for hormone and trace element concentrations; eight were studied for microbial contamination. The concentrations of CRH, hPL, oxytocin and ACTH in samples of raw, steamed dehydrated and raw dehydrated placental tissue were detected using ELISA. A yeast bioassay was performed in order to detect estrogen equivalent (EEQ) and gestagen equivalent (PEQ) active substances. Elements (As, Cd, Fe, Pb, Se, Hg) were analyzed using ICP-MS. Isolated colonies from tissue and placenta swab samples were identified using Vitek MS.

Results: Following mean hormone concentrations were detected in raw placental tissue: CRH (177.88 ng/g), hPL (17.99 mg/g), oxytocin (85.10 pg/g), ACTH (2.07 ng/g), estrogen equivalent active substances (46.95 ng/g) and gestagen equivalent active substances (2.12 μg/g). All hormones were sensitive to processing with a significant concentration reduction through steaming and dehydration. Microorganisms mainly from the vaginal flora were detected on placenta swab samples and samples from raw, steamed, dehydrated and steamed dehydrated tissue and mostly disappeared after dehydration. According to regulations of the European Union the concentrations of potentially toxic elements (As, Cd, Hg, Pb) were below the toxicity threshold for foodstuffs.

Conclusion: The commonly used protocols for preparation of placenta for its individual oral ingestion reduce hormone concentrations and bacterial contamination.
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http://dx.doi.org/10.1016/j.placenta.2018.05.006DOI Listing
July 2018

Comparison of sample preparation techniques and data analysis for the LC-MS/MS-based identification of proteins in human follicular fluid.

Am J Reprod Immunol 2018 08 25;80(2):e12994. Epub 2018 Jun 25.

Host Septomics Research Group, Jena University Hospital, Jena, Germany.

The proteomic analysis of complex body fluids by liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis requires the selection of suitable sample preparation techniques and optimal parameter settings in data analysis software packages to obtain reliable results. Proteomic analysis of follicular fluid, as a representative of a complex body fluid similar to serum or plasma, is difficult as it contains a vast amount of high abundant proteins and a variety of proteins with different concentrations. However, the accessibility of this complex body fluid for LC-MS/MS analysis is an opportunity to gain insights into the status, the composition of fertility-relevant proteins including immunological factors or for the discovery of new diagnostic and prognostic markers for, for example, the treatment of infertility. In this study, we compared different sample preparation methods (FASP, eFASP and in-solution digestion) and three different data analysis software packages (Proteome Discoverer with SEQUEST, Mascot and MaxQuant with Andromeda) combined with semi- and full-tryptic databank search options to obtain a maximum coverage of the follicular fluid proteome. We found that the most comprehensive proteome coverage is achieved by the eFASP sample preparation method using SDS in the initial denaturing step and the SEQUEST-based semi-tryptic data analysis. In conclusion, we have developed a fractionation-free methodical workflow for in depth LC-MS/MS-based analysis for the standardized investigation of human follicle fluid as an important representative of a complex body fluid. Taken together, we were able to identify a total of 1392 proteins in follicular fluid.
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http://dx.doi.org/10.1111/aji.12994DOI Listing
August 2018

Breast carcinoma in pregnancy with spheroid-like placental metastases-a case report.

APMIS 2018 May 17;126(5):448-452. Epub 2018 Apr 17.

Department of Pathology, Center for Pediatric and Pregnancy Related Pathology, Oslo University Hospital, Oslo, Norway.

Breast cancer is one of the most common malignancies diagnosed in pregnancy. Although the tumor is often detected at an advanced stage, placental metastases are rare. Here, we describe the case of a woman with breast cancer recurrence during pregnancy and subsequent metastases. The focus of this study is the large amount of placenta metastases, which have been analyzed immunohistochemically. Staining with trophoblast markers (placenta alkaline phosphatase, beta human chorionic gonadotropin and human placental lactogen) showed the strict localization of metastases in the intervillous space without invasion into fetal tissue. They have a large spheroidal shape and are free of blood vessels. Staining with Ki-67 revealed an outer proliferative shell and inner necrotic core. At week 28, a healthy newborn was born by elective cesarean section. A few weeks later, after surgery and FEC60 (fluorouracil, epirubicin, cyclophosphamide) cycles, the patient died. Breast cancer metastases in the placenta are rarely described. The special immunological environment in pregnancy may influence phenotype, growth, and behavior of tumor and metastases.
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http://dx.doi.org/10.1111/apm.12827DOI Listing
May 2018
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