Dm Baston-Büst

Syndecan-1 knock-down in decidualized human endometrial stromal cells leads to significant changes in cytokine and angiogenic factor expression patterns

BackgroundSuccessful embryonic implantation depends on a synchronized embryo-maternal dialogue. Chemokines, such as chemokine ligand 1 (CXCL1), play essential roles in the maternal reproductive tract leading to morphological changes during decidualization, mediating maternal acceptance towards the semi-allograft embryo and induction of angiogenesis. Chemokine binding to their classical G-protein coupled receptors is essentially supported by the syndecan (Sdc) family of heparan sulfate proteoglycans. The aim of this study was to identify the involvement of Sdc-1 at the embryo-maternal interface regarding changes of the chemokine and angiogenic profile of the decidua during the process of decidualization and implantation in human endometrium.MethodsA stable Sdc-1 knock-down was generated in the immortalized human endometrial stromal cell line St-T1 and was named KdS1. The ability of KdS1 to decidualize was proven by Insulin-like growth factor binding 1 (IGFBP1) and prolactin (PRL) confirmation on mRNA level before further experiments were carried out. Dot blot protein analyses of decidualized knock-down cells vs non-transfected controls were performed. In order to imitate embryonic implantation, decidualized KdS1 were then incubated with IL-1beta, an embryo secretion product, vs controls. Statistical analyses were performed applying the Students t-test with p < 0.05, p < 0.02 and p < 0.01 and one way post-hoc ANOVA test with p < 0.05 as cut-offs for statistical significance.ResultsThe induction of the Sdc-1 knock-down revealed significant changes in cytokine and angiogenic factor expression profiles of dKdS1 vs decidualized controls. Incubation with embryonic IL-1beta altered the expression patterns of KdS1 chemokines and angiogenic factors towards inflammatory-associated molecules and factors involved in matrix regulation.ConclusionsSdc-1 knock-down in human endometrial stroma cells led to fulminant changes regarding cytokine and angiogenic factor expression profiles upon decidualization and imitation of embryonic contact. Sdc-1 appears to play an important role as a co-receptor and storage factor for many cytokines and angiogenic factors during decidualization and implantation period, supporting proper implantation and angiogenesis by regulation of chemokine and angiogenic factor secretion in favour of the implanting embryo.

CXCR7 and syndecan-4 are potential receptors for CXCL12 in human cytotrophoblasts

The placenta forms the interface between the mother and the fetus. During placental development cytotrophoblasts differentiate to form the syncytium or to invade the decidual wall to breach maternal vessels and establish the blood flow in the intervillous space. This process is still not well understood but it is proposed that chemokines and their receptors are involved in guiding cytotrophoblasts to the decidua and maternal vessels as well as attracting immunocompetent cells to the implantation site. CXCL12 is a chemokine expressed by cytotrophoblasts and is involved in cytotrophoblast invasion, differentiation and survival. One of its receptors, CXCR4, has been detected on cytotrophoblasts. Recent data show that CXCR7 and syndecan-4 might partially mediate CXCL12 function in other cell types. In this study, we examined CXCR7 and syndecan-4 expression at the maternal-fetal interface via immmunolocalization in placental tissue sections and in isolated cytotrophoblasts. We further used immunoblot analyses to confirm the data. We were able to show that cytotrophoblasts express both receptors and that upregulation occurs during the differentiation process of cytotrophoblasts towards the invasive phenotype. On a functional level CXCR7 seems not to be involved in JAR cell chemotaxis, suggesting a different function of this receptor. In conclusion, we propose that CXCL12 binds to CXCR4, but also to CXCR7 and syndecan-4. These three receptors could mediate different functions of CXCL12, such as cell migration, directed invasion, proliferation and survival. The latter molecules might also be involved in the development of placental pathologies, such as preeclampsia or choriocarcinoma growth.

hCG stimulates angiogenic signals in lymphatic endothelial and circulating angiogenic cells

Human chorionic gonadotropin (hCG) has long been associated with the initiation and maintenance of pregnancy, where angiogenesis plays an important role. However, the function of hCG in angiogenesis and the recruitment of vascular active cells are not fully understood. In this study, the role of hCG and its receptor in circulating angiogenic and human endothelial cells, including lymphatic, uterine microvascular, and umbilical vein endothelial cells, was examined. Immunohistochemistry and immunoblot analysis were used to detect LH/hCG receptor expression and the expression of hCG-induced angiogenic molecules. HIF-1α was determined via ELISA and downstream molecules, such as CXCL12 and CXCR4, via real-time PCR. Chemotaxis was analyzed using Boyden chambers. Our results show that the LH/hCG receptor was present in all tested cells. Furthermore, hCG was able to stimulate LH/hCG-receptor-specific migration in a dose-dependent fashion and induce key angiogenic molecules, including HIF-1α, CXCL12, and CXCR4. In conclusion, our findings underscore the importance of hCG as one of the first angiogenic molecules produced by the conceptus. hCG itself alters endothelial motility, recruitment, and expression of pro-angiogenic molecules and may therefore play an important role in vascular adaption during implantation and early placental formation.

A possible ambivalent role for relaxin in human myometrial and decidual cells in vitro.

PurposeBased on the reported tocolytic action of the hormone relaxin (RLX) in rodents, locally produced in reproductive tissues and the corpus luteum in mammals, the present study aimed to evaluate the influence of RLX on contraction-mediating cyclooxygenases-1 and -2 (COX) and the contractile prostaglandin PGE2 in human myometrial and decidual cells. Primary cultured cells were obtained from uteri and placentas of term and preterm women undergoing elective caesarean section.MethodsIn vitro culture of primary myometrial and decidual cells, immunocytochemistry, reverse transcription and real-time PCR, Western blot, ELISA.ResultsWe demonstrate for the first time an activating effect of RLX for human COX-1 and COX-2 in primary myometrial and decidual cells in vitro.ConclusionsThese effects might potentially contribute to birth-associated induction of contractions in vivo.

Wann ist die Chance auf Mutterschaft vertan

Viele Frauen, die die Familienplanung immer weiter hinausschieben, verdrängen die Tatsache, dass ihre Fertilität etwa ab dem 35. Lebensjahr drastisch sinkt. Was heute über die „Biologie des reproduktiven Alters“ bekannt ist und mit welchen Methoden Frauen im kritischen Alter doch noch zu einem eigenen Kind verholfen werden kann, ist Thema des folgenden Beitrags.

S2k-Leitlinie: Fertilitätserhaltende Maßnahmen bei onkologischen Erkrankungen

ZusammenfassungOnkologische und nichtonkologische Erkrankungen können die gegenwärtige oder zukünftige Fertilität beeinträchtigen, entweder durch die Krankheit selbst oder durch notwendig chirurgische Eingriffe, hormonelle und/oder gonadotoxische Behandlungen. Sie erfordern damit einen adäquaten fertilitätsprotektiven Ansatz – sprich eine unbedingte Beratung und, wenn die Voraussetzungen gegeben sind, auch eine entsprechende fertilitätsprotektive Therapie. Die Kryokonservierung von Oozyten stellt zumeist die Methode der ersten Wahl bei postpubertären Frauen dar. Metaphase-II-Oozyten-Kryokonservierung mittels Vitrifikation ist dabei die bevorzugte und empfohlene Option. Aber auch der kumulative Nachweis der Wiederherstellung der endokrinen Ovarialfunktion und der daraus resultierenden spontanen bzw. nach orthotoper Transplantation von kryokonserviertem Ovarialgewebe und Einsatz unterstützender assistierter reproduktionsmedizinischer Techniken resultierenden Schwangerschaften und Geburten spricht für diese Methode als offene klinische Anwendung – insbesondere bei präpubertären Mädchen, die nicht minder von keimzellschädigenden Behandlungen betroffen sind als adoleszente oder adulte Patientinnen im fertilen Alter und bei denen keine andere nachweislich wirksame Option zum Fertilitätserhalt derzeit verfügbar ist.AbstractOncological and nononcological diseases can affect current or future fertility, either through the disease itself or through necessary surgery, hormonal and/or gonadotoxic treatments. They therefore require an adequate fertility-protective approach—i. e. counselling is essential and, if the prerequisites are met, an appropriate fertility-protective therapy. Cryopreservation of oocytes is usually the first-choice method for postpubertal women. Metaphase II oocyte cryopreservation by vitrification is the preferred and recommended option. However, cumulative evidence of restoration of ovarian endocrine function and resulting spontaneous pregnancies and births, or after the use of assisted reproductive techniques, after orthotopic transplantation of cryopreserved ovarian tissue also advocates this method as an open clinical application, especially in prepubertal girls who are no less affected by germ cell-damaging treatments than adolescent or adult patients of reproductive age and for whom no other possible option for fertility preservation is currently available.

Concept Paper on the Technique of Cryopreservation, Removal and Transplantation of Ovarian Tissue for Fertility Preservation

The cryopreservation of ovarian tissue with subsequent transplantation of the tissue represents an established method of fertility protection for female patients who have to undergo gonadotoxic therapy. The procedure can be performed at any point in the cycle and thus generally does not lead to any delay in oncological therapy. With the aid of this procedure, more than 130 births to date worldwide have been able to be recorded. The birth rate is currently approximately 30% and it can be assumed that this will increase through the further optimisation of the cryopreservation and surgical technique. The concept paper presented here is intended to provide guidance for managing cryopreservation and transplantation of ovarian tissue to German-speaking reproductive medicine centres.

Rettung vor der Infertilität im Alter

Über Social Freezing, dem vorsorglichen Einfrieren von Eizellen, damit eine Frau auch noch in späteren Jahren schwanger werden kann, wird viel diskutiert. Im Mittelpunkt des nachfolgenden Beitrags stehen weniger ethische als vielmehr medizinische und gesellschaftliche Aspekte. Wie funktioniert Sozial Freezing, welche Erfolgsraten sind zu erwarten, wer lässt sich behandeln, und was kostet die Behandlung?