Embryonic Implantation Research Articles

Effect of Blood Gel Derivatives on Wound Healing in Mouse Injured Tissue Models.

Several previous studies in the field of assisted reproduction have focused on the use of blood gel derivatives, such as platelet-rich fibrin (PRF), as a treatment for endometrial rehabilitation. However, the ability to release growth factors and the gel form of this product led to the evolution of platelet lysates. In this study, blood gel derivatives, including PRF lysate, which was in liquid form, and PRF gel, were collected and evaluated for growth factors. It was shown to be effective in endometrial wound healing and regeneration in mouse injured uterine tissue models through structure and function (pinopode expression, embryo implantation) evaluation. The results demonstrated that the concentrations of growth factors, including PDGF-AB and VEGF-A, were higher in the PRF lysate compared to the PRF gel (p < 0.05). PRF lysate could release these growth factors for 8 days. Furthermore, both PRF gel and PRF lysate restored the morphology of injured endometrial tissues in terms of luminal and glandular epithelia, as well as uterine gland secretory activity. However, the presence of pinopodes and embryonic implantation were only observed in the PRF lysate group. It can be concluded that PRF lysate promotes wound healing in mouse injured tissue models in vitro, which can act as healing products in tissue repair.

Immune Regulation of Seminal Plasma on the Endometrial Microenvironment: Physiological and Pathological Conditions.

Seminal plasma (SP) accounts for more than 90% of semen volume. It induces inflammation, regulates immune tolerance, and facilitates embryonic development and implantation in the female reproductive tract. In the physiological state, SP promotes endometrial decidualization and causes changes in immune cells such as macrophages, natural killer cells, regulatory T cells, and dendritic cells. This leads to the secretion of cytokines and chemokines and also results in the alteration of miRNA profiles and the expression of genes related to endometrial tolerance and angiogenesis. Together, these changes modulate the endometrial immune microenvironment and contribute to implantation and pregnancy. However, in pathological situations, abnormal alterations in SP due to advanced age or poor diet in men can interfere with a woman's immune adaptation to pregnancy, negatively affecting embryo implantation and even the health of the offspring. Uterine pathologies such as endometriosis and endometritis can cause the endometrium to respond negatively to SP, which can further contribute to pathological progress and interfere with conception. The research on the mechanism of SP in the endometrium is conducive to the development of new targets for intervention to improve reproductive outcomes and may also provide new ideas for semen-assisted treatment of clinical infertility.

Analysis of potential copy-number variations and genes associated with first-trimester missed abortion

BackgroundCopy number variation sequencing (CNV-seq) was proven to be a highly effective tool in studying of chromosomal copy number variations (CNVs) in prenatal diagnosis and post-natal cases with developmental abnormalities. However, the overall characteristics of missed abortion (MA) CNVs were largely unexplored. MethodsWe retrospectively analyzed the results of CNV-seq in first-trimester MA. The samples included were single pregnancy loss before 13 gestational weeks, and other potential factors affecting embryonic implantation and development had been excluded. Gene ontology and KEGG enrichment analysis was performed on the smallest overlapping regions (SORs) of high-frequency deletion/duplication. ResultOn the basis of strict inclusion and exclusion criteria, only 152 samples were included in our study. 77 (50.7%) samples displayed chromosome number abnormalities, 32 (21%) showed isolated CNVs, and 43 (28.3%) showed no CNVs. A total of 45 CNVs, ranging in size between 300 Kb and 126.56 Mb were identified, comprising 13 segmental aneuploidies CNVs, and 32 submicroscopic CNVs. Among these CNVs, we screened out four SORs (5q31.3, 5p15.33-p15.2, 8p23.3-p23.2, and 8q22.2–24.3), which were potentially associated with first-term MA. 16 genes were identified as potential miscarriage candidate genes through gene-prioritization analysis, including three genes (MYOM2, SDHA and TPPP) critical for embryonic heart or brain development. ConclusionWe identified some potential candidate CNVs and genes associated with first-trimester MA. 5q31.3 duplications, 5p15.33-p15.2 deletions, 8p23.3-p23.2 deletions and 8p22.2-p24.3 duplications are four potential candidate CNVs. Additionally, MYOM2, SDHA and TPPP are potential genes associated with first-trimester MA.

P-472 Endometrial stimulation with culture medium containing granulocyte macrophage colony-stimulating factor promotes embryo implantation after vitrified-warmed blastocyst transfer

Abstract Study question Does endometrial stimulation with culture medium containing granulocyte macrophage colony-stimulating factor (GM-CSF) promote implantation potential of vitrified-warmed blastocysts? Summary answer Endometrial stimulation with culture medium containing GM-CSF prior to transfer of vitrified-warmed blastocysts may promote embryo implantation and improve pregnancy continuation. What is known already Human embryonic development and implantation in vivo is precisely regulated by various cytokines, including growth factors. GM-CSF is a cytokine that plays an important role in reproductive function. We reported the possibility that the use of culture medium containing GM-CSF in cleavage stage vitrified-warmed embryo transfers (ETs) promotes embryo implantation (ESHRE2018). However, it has been reported that endometrial stimulation with culture supernatant up to blastocyst stage (SEET method) improves embryo implantation, but the effect of endometrial stimulation with culture medium containing GM-CSF (G-SEET method) on embryo implantation is not clear. Study design, size, duration This study was conducted at a single in vitro fertilization (IVF) center from June 2020 to February 2022. The G-SEET procedure was performed at each patient’s request. All study participants provided informed consent, and the study design was approved by the ethics committee of IVF Nagata Clinic, Fukuoka, Japan. Participants/materials, setting, methods We examined 702 cycles of vitrified-warmed blastocyst transfer. Warmed blastocysts were incubated in recovery culture for several hours and transferred on day 5 (control group). For the G-SEET method, only culture medium containing GM-CSF was injected into the uterus two days before ET, which was performed as in the control group (G-SEET group). The human chorionic gonadotropin (hCG) positive, clinical pregnancy, miscarriage, and ongoing pregnancy rates were compared between the two groups. Main results and the role of chance Of the 702 cycles of single vitrified-warmed blastocyst transfers, the control and G-SEET groups had 459 and 243 cycles, respectively. The G-SEET group had a higher patient age (37.5±4.2 vs. 36.2±3.8, p &amp;lt; 0.01), a thinner endometrial thickness (10.3±1.9 vs. 10.9±2.0, p &amp;lt; 0.01), a lower percentage of good blastocyst transfers (65.4% vs. 79.5%, p &amp;lt; 0.01), and a higher cumulative number of implantation failures (3.6±2.5 vs. 1.9±1.9, p &amp;lt; 0.01) than the control group. The G-SEET method was performed at the patient’s request, so the number of cycles and patient background varied. Multivariate analysis was performed using female age, male age, number of miscarriages, endometrial thickness, presence of good blastocysts, and cumulative number of implantation failures as confounding factors. Compared with the control group, the G-SEET group exhibited the following: hCG positivity rate [odds ratio (OR): 1.14, 95% confidence interval (CI): 0.79–1.64, p = 0.481], clinical pregnancy rate (OR: 1.81, 95% CI: 1.21–2.71, p = 0.003), miscarriage rate (OR: 0.735, 95% CI: 0.30–1.76, p = 0.491), and ongoing pregnancy rate (OR: 1.79, 95% CI: 1.20–2.69, p = 0.004). Clinical pregnancy and ongoing pregnancy rates were significantly higher in the G-SEET group compared with the control group. Limitations, reasons for caution The study was limited by the study size and lack of data about live birth rates after ET. In addition, intrauterine infusion of GM-CSF-free culture medium should be used as a control, but this was difficult in the private clinic setting. Wider implications of the findings This study suggests that endometrial stimulation with culture media containing GM-CSF prior to transfer of vitrified-warmed blastocysts promotes embryo implantation and enhances the ongoing pregnancy. The G-SEET method may be clinically useful as a new ET method. Trial registration number not applicable

P-480 An endometrial receptivity scoring system evaluated by ultrasonography in patients undergoing frozen-thawed embryo transfer: a prospective cohort study

Abstract Study question Can a scoring system based on ultrasound indicators evaluate the endometrial receptivity (ER) in a noninvasive and effective manner? Summary answer A noninvasive ultrasound scoring system was proposed, through which, a patient’s ER situation can be assessed in a noninvasive, efficient and accurate manner. What is known already Endometrial receptivity (ER) refers to the receptiveness of the endometrium to embryonic implantation and subsequent development. The endometrium plays a crucial role in embryo implantation, especially when the embryo quality is good. Ultrasound technology has become a routine method for ER evaluation. The independent evaluation value of various ultrasonic indicators is controversial. Some researchers designed a multi-index prediction system, but the prediction results vary. To further understand ER, we performed this prospective cohort study to help evaluate ER in a noninvasive and efficient manner. Study design, size, duration This prospective study included 197 infertile women undergoing FET from April 2019 to July 2021. Transvaginal three-dimensional ultrasound was performed on the transfer day to evaluate ER, including endometrial thickness, morphology, volume, movement, blood flow and flow index. Participants/materials, setting, methods This study was conducted at the Reproductive and Genetic Hospital of CITIC-Xiangya. The primary outcome of this study was the clinical pregnancy rate. All included patients were divided into a pregnant group and a nonpregnant group according to whether clinical pregnancy was achieved. Main results and the role of chance A total of 197 FET patients with 139 pregnancies (70.5%) were analysed. Primary infertility (adjusted odds ratio (aOR), 1.98; 95% confidence interval (CI), 1.01–3.882; P = 0.047) and more frequent endometrial peristalsis (aOR, 1.33; 95% CI, 1.028–1.722; P = 0.03) were protective factors against clinical pregnancy. Scores of 1 to 2 were given according to the relationship between different groups of ultrasonic indices and the clinical pregnancy rate (CPR). A patient’s ER score was the sum of the scores of the 6 items. The ER score was significantly higher in the pregnant group than that in the nonpregnant group (7.40 ± 1.73 versus 6.33 ± 1.99, P = 0.001). The CPR increased as the ER score increased; the CPR significantly improved for the group with total ER scores of &amp;lt; 6 to ≥ 6 (45.5% versus 75.6%, P = 0.001). Limitations, reasons for caution First, this study only included natural cycles of FET. The application for other populations remains to be verified. Second, perhaps with the progress of ultrasonic technology, there will be new and more direct indicators. Third, we did not further explore the relationship between ER and the live birth rate. Wider implications of the findings The scoring system provides a broader idea for guiding clinical practice to perform more efficient transplantation and provide a noninvasive evaluation of ER in the future. Trial registration number None

P-803 3D human endometrium from noninvasively retrieved primary endometrial cells is a reliable model to mimic the implantation window

Abstract Study question Can human endometrial organoids (hEOs) from a noninvasive source (menstrual blood-MB) mimic the physiology of a receptive endometrium Summary answer Endometrial organoids from MB functionally respond to stimuli and express markers of receptivity and can be modulated. What is known already 3D models are considered a new step in promoting precision medicine and an advanced tool for studying endometrial biology, endometrial-associated diseases, and understanding the complex mechanisms that comprise endometrial-embryo crosstalk. So far, both 2D and 3D models are usually obtained from endometrial biopsy. The recent discovery of molecules crucial for successful embryo implantation has offered researchers valuable insights in this field. However, important questions about the molecular mechanisms that drive this process remain to be deciphered. Study design, size, duration hEOs are usually obtained from endometrial biopsy, in this case a noninvasive source(MB) was used to isolate endometrial epithelial (Ep) and stromal (hESC) cells. The hEOs were generated to study the morphological changes that occur in vivo during the menstrual cycle, particularly in the window of implantation (WOI). The hEOs obtained from MB were exposed to hormones treatments to mimic WOI and were also treated with the progesterone blocker mifepristone to reverse the decidualization process. Participants/materials, setting, methods 3D hEOs were prepared from 6 healthy volunteers and cultured for 4 days in expansion medium supplemented with 10-8 M E2 + 10-6 M P4 and 500 uM cAMP. As a control, hEOs were treated with the progesterone blocker mifepristone to reverse the decidualization process. Finally, hEOs were washed with PBS and fixed for immunofluorescent staining or for scanning-electron-microscopy (SEM) or stored at -80 °C for gene expression analysis. Main results and the role of chance In this study, we were able to establish a reliable 3D model using both epithelial and stromal cells with a less invasive approach to mimic the mid-secretory phase. Immunofluorescence staining of vimentin (marker of stromal cells) and cytokeratin 19 (marker of epithelial cells) confirmed that organoids are formed by both Ep and hESCs. Using SEM, we closely observed the luminal surface of hEOs in proliferative phase, where the inner side of Ep cells was characterized by the presence of pinopods, a specific marker of the implantation window. In addition, hEOs express PP14, an implantation marker that has been shown to be significantly up-regulated during WOI. Finally, the expression of FKBP5, regulated by cAMP, and ZBP16, regulated by progesterone, increased significantly during decidualization, while their expression was up-regulated at dawn by mifepristone, a progesterone blocker, thus confirming that hEOs are an effective model of WOI. Limitations, reasons for caution All results need to be validated in a larger cohort and further implemented with other cell types (e.g., endothelial and immune cells) to implement this model for drug testing and therapy Wider implications of the findings Effective and reliable modeling of embryonic implantation is needed to mimic the cascade of molecular events that occurs in vivo. This model can be used for personalized medicine without the invasiveness of biopsy. Trial registration number Not applicable

The Extracellular Vesicles: Their Role and Future in Reproduction and Embryonic Implantation

Intercellular communication is an integral part of the physiological process in a living organism. Evidence has shown that a significant amount of human protein-coding genes is endowed with the production of membrane and secreted protein types. Some of these proteins possessed post-translational influence that interferes with cellular functions. Recently the concept of cell communication gravitated to the use of membrane-bound particles called Extracellular Vesicles (EVs). EVs are released from the host cell and impact on the recipient cell through its contents (Cargo). The cargo contains various particles such as lipids, proteins, RNA, and glycolipids derived from the host cells. The current widespread research in EVs has brought its mediating role in physiological and pathological processes to limelight. Though the regulatory part of EVs has been widely embraced by researchers, the discrepancy associated with its collection and isolation as well as its analysis has remained a subject of debate in the literature. The facts emanating from the literature suggest that EVs, with its prospective application in Reproductive biology, has prompted extensive research in the last decade. Its knowledge has broadened the scope of understanding the physiology and pathology scenarios involved in reproductive processes. As a potential biomarker, it has become a valuable tool for diagnosis, prognostic, and therapeutic purposes, especially in the context of reproductive processes. However, the challenges of standardizing the isolation, purification, and analyzing EVs have remained a nightmare that need to be surmounted. Emerging evidence has demonstrated its impact in gamete development, fertilization, and embryo implantation. Thus, it could serve as a platform to understand the mechanism of conception and implantation. By extension, define a therapeutic approach for women with recurrent pregnancy loss.

Time-Restricted Feeding Ameliorates Uterine Epithelial Estrogen Receptor α Transcriptional Activity at the Time of Embryo Implantation in Mice Fed a High-Fat Diet

BackgroundMore than 30% of reproductive-age women are obese or overweight. Obesity and exposure to a high-fat diet (HFD) detrimentally affect endometrial development and embryo implantation. We previously reported that time-restricted feeding (TRF) improved ovarian follicular development, but whether and how TRF modulates embryo implantation are poorly understood. ObjectiveWe investigated the effect of TRF on embryo implantation. MethodsIn TRF group, mice had 10 h of food free access from 9 pm to 7 am, and fed a normal diet or a HFD. Tail vein injection of Chicago blue dye was used to examine embryo implantation sites at day 5.5 (D5.5) of pregnancy. Serum collected at D0.5 and D4.5 of pregnancy was used to examine the level of estradiol (E2) and progesterone. Uterine estrogen receptor (ER) and progesterone receptor levels and their targeted aquaporins (AQPs) were measured. LC–MS was used to analyze bile acid (BA) composition, and primary hepatocytes were used to test the effects of BA on the expression level of SULT1E1, a key enzyme in estrogen inactivation and elimination. ResultsWe found that TRF prevented HFD-induced embryo loss and alleviated the defect in luminal closure on D4.5 of pregnancy. The cyclic changes of E2 level were lost in mice fed ad libitum but not in TRF mice on the HFD. The HFD increased ER-α expression and transcriptional activity, which induced AQP3 and AQP5 expression on D4.5 of pregnancy. TRF prevented the negative effect of the HFD on uterine luminal closure. Furthermore, in vitro and in vivo results showed that BA suppressed estrogen degradation by activating liver SULT1E1 expression. ConclusionsOur findings demonstrated that TRF prevented HFD-induced defects in luminal closure, thereby improving embryonic implantation, and provide novel insights into the effects of dietary intervention on obesity and associated infertility.

Rhythm of the First Language: Dynamics of Extracellular Vesicle-Based Embryo-Maternal Communication in the Pre-Implantation Microenvironment.

One of the most critical steps in mammalian reproduction is implantation. Embryos with an impaired capacity for embryo-maternal crosstalk are thought to have a reduced potential for implantation. One agent of embryo-maternal communication is extracellular vesicles (EV). EVs are lipid bilayer-bound biological nanoparticles implicated in intercellular communication between many of the known cell types. In the current study, we isolated EVs from trophoblast analogue JAr spheroids and supplemented the EVs with receptive endometrium analogue RL95-2 cells to simulate pre-implantation embryo-maternal dialogue. The transcriptome of the endometrial cells was examined at 30 min, 4 h and 48 h intervals using Oxford Nanopore® technology. At the time points, 30 min, 4 h and 48 h, the endometrial cells showed a significantly altered transcriptome. It seems trophoblast EVs induce a swift and drastic effect on the endometrial transcriptome. The effect peaks at around 4 h of EV supplementation, indicating a generalized effect on cell physiology. Alterations are especially apparent in biological pathways critical to embryonic implantation, such as extracellular matrix-receptor interactions and cytokine-receptor interactions. These observations can be helpful in elucidating the dynamics of embryo-maternal communication in the pre-implantation period.

Exosomes: New regulators of reproductive development.

Exosomes are a subtype of extracellular vesicles (EVs) with a size range between 30 and 150​nm, which can be released by the majority of cell types and circulate in body fluid. They function as a long-distance cell-to-cell communication mechanism that modulates the gene expression profile and fate of target cells. Increasing evidence has indicated exosomes' central role in regulating various complex reproductive processes. However, to our knowledge, a review that focally and vividly describes the role of exosomes in reproductive development is still lacking. This review highlights our knowledge about the contribution of exosomes to early mammalian reproduction, such as gametogenesis, fertilization, early embryonic development, implantation, placentation and pregnancy. The discussion is primarily drawn from literature pertaining to the mammalian lineage with emphasis on the roles of exosomes in human reproduction and laboratory and livestock models.

Failure to Connect: Experimental Evidence that Benzo(a)pyrene Impedes Embryonic Implantation.

Failure to Connect: Experimental Evidence that Benzo(a)pyrene Impedes Embryonic Implantation.

Exploration of Maternal and Fetal Toxicity Risks for Metronidazole-Related Teratogenicity and Hepatotoxicity through an Assessment in Albino Rats.

Metronidazole is the primary antimicrobial drug for treating acute and chronic vaginal pathogens during pregnancy; however, there has been insufficient research on placental disorders, early pregnancy loss, and preterm birth. Here, the potential activity of metronidazole on pregnancy outcomes was investigated. 130 mg/kg body weight of metronidazole was orally given individually to pregnant rats on gestation days 0-7, 7-14, and 0-20. Pregnancy outcome evaluations were carried out on gestation day 20. It was demonstrated that metronidazole could induce maternal and fetal hepatotoxicity. There is a significant increase in the activities of maternal hepatic enzymes (ALT, AST, and ALP), total cholesterol, and triglycerides compared with the control. These biochemical findings were evidenced by maternal and fetal liver histopathological alterations. Furthermore, metronidazole caused a significant decrease in the number of implantation sites and fetal viability, whereas it caused an increase in fetal lethality and the number of fetal resorptions. In addition, a significant decrease in fetal weight, placental weight, and placental diameter was estimated. Macroscopical examination revealed placental discoloration and hypotrophy in the labyrinth zone and the degeneration of the basal zone. The fetal defects are related to exencephaly, visceral hernias, and tail defects. These findings suggest that the administration of metroniazole during gestation interferes with embryonic implantation and fetal organogenesis and enhances placental pathology. We can also conclude that metronidazole has potential maternal and fetal risks and is unsafe during pregnancy. Additionally, it should be strictly advised and prescribed, and further consideration should be given to the associated health risks.

Exosomes: The role in mammalian reproductive regulation and pregnancy-related diseases.

Exosomes are a kind of extracellular vesicles that are produced and secreted by different mammalian cells. They serve as cargo proteins and can transfer different kinds of biomolecules, including proteins, lipids, and nucleic acids, which consequently act on target cells to exert different biological effects. Recent years have witnessed a significant increase in the number of studies on exosomes due to the potential effects of exosomes in the diagnosis and treatment of cancers, neurodegenerative diseases, and immune disorders. Previous studies have demonstrated that exosomal contents, especially miRNAs, are implicated in numerous physiological processes such as reproduction, and are crucial regulators of mammalian reproduction and pregnancy-related diseases. Here, we describe the origin, composition, and intercellular communication of exosomes, and discuss their functions in follicular development, early embryonic development, embryonic implantation, male reproduction and development of pregnancy-related diseases in humans and animals. We believe this study will provide a foundation for revealing the mechanism of exosomes in regulating mammalian reproduction, and providing new approaches and ideas for the diagnosis and treatment of pregnancy-related diseases.

Endometrial Injury Upregulates Expression of Receptivity Genes in Women with Implantation Failure.

Homeobox genes A10 (HOXA10) and A11 (HOXA11), members of the abdominal B gene family, are responsible for embryonic survival and implantation. This study was planned to investigate whether endometrial injury alters the expression of both transcripts in women with implantation failure. A total of 54 women with implantation failure were divided into two equal groups as experimental (scratching) and sham (no scratching). Participants in the scratching group were exposed to endometrial injury in the mid-luteal phase, and those in the sham group were exposed to endometrial flushing. The scratching group, but not the sham group, underwent prior endometrial sampling. A second endometrial sampling was performed on the scratching group in the mid-luteal phase of the following cycle. The mRNA and protein levels of the HOXA10 and 11 transcripts were determined in endometrial samples collected before and after injury/flushing. Participants in each group underwent IVF/ET in the cycle after the second endometrial sampling. Endometrial injury caused a 60.1-fold (p < 0.01) increase in HOXA10 mRNA and a 9.0-fold increase in HOXA11 mRNA (p < 0.02). Injury resulted in a significant increase in both HOXA10 (p < 0.001) and HOXA11 protein expression (p < 0.003). There was no significant change in HOXA10 and 11 mRNA expressions after flushing. Clinical pregnancy, live birth, and miscarriage rates of the both groups were similar. Endometrial injury increases homeobox transcript expression at both mRNA and protein levels.

Screening genes related to embryo implantation in Dazu black goats (Capra Hircus) by morphological and transcriptome analyses.

Embryo implantation is a critical step in the establishment of pregnancy. However, the mechanisms of embryo implantation during early pregnancy in goats remain unclear due to the lack of published studies examining the genes involved in embryo implantation. As a popular goat breed in southwest China, Dazu black goats (DBGs) are highly adaptable and exhibit high fertility, making this breed a good model in which to study reproductive performance of goats. Here, morphological analysis showed that compared with the non-pregnant (NP) groups, the endometrial thickness of the goats in the P15 and P19 groups (15 and 19-day pregnant groups, respectively) were increased (P < 0.01). Proliferating Cell Nuclear Antigen (PCNA) staining showed that PCNA was expressed in the NP, P15, and P19 groups. Transcriptome analysis was then conducted to identify gene expression patterns in uterine tissue during DBG embryo implantation. By comparing uterine tissue at different stages of embryonic implantation, 48 in NP_vs._P15, 318 in NP_vs._P19, and 1439 in P15_vs._P19, differentially expressed mRNAs were identified. Gene Ontology (GO) enrichments of the differentially expressed genes were enriched in the extracellular region, extracellular space, transporter activity, extracellular region, immune system process, immune response, and defense response etc. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, the biological metabolic pathways with which the differentially expressed genes are associated were explored. Through KEGG analysis, the DBGs were associated with oxidative phosphorylation, complement and coagulation cascades, arginine and proline metabolism, metabolic pathways, arachidonic acid metabolism, and ECM-receptor interaction. These candidate genes (CSF1, C1S, CST6, SLC24A4, HOXA10, HOXA11, MMP9, and ITGA11) and enriched signaling pathways could be valuable references for exploring the molecular mechanisms underlying goat embryo implantation.

Endometrial Status and Embryonic Implantation about 150 Cases at the Medically Assisted Procreation Laboratory “Le Diafounou”

Endometrial Status and Embryonic Implantation about 150 Cases at the Medically Assisted Procreation Laboratory “Le Diafounou”

Implantation of Embryonic Cardiomyocytes in a Post-Infarction Myocardium: A Long-Term Outcome after 25 Years of Follow-Up.

In this article, we report the first case of embryonic cardiomyocyte implantation in a patient with severe ischemic heart lesions. In 1998, a 58-year-old male underwent coronary artery bypass grafting supplemented by injection of a pre-prepared suspension of human embryonic cardiomyocytes. During the next 24 years, the patient felt well and reported quite satisfactory quality of life. He is currently 82 years old and also feels well. He has no signs of circulatory insufficiency. No complications associated with cell implantation were observed during the entire follow-up period.

Aquaporin 7 is upregulated through the PI3K-Akt pathway and modulates decidualisation of endometrial stromal cells.

Aquaporin 7 (AQP7) is selectively expressed in decidualised endometrial stromal cells (ESCs) of mice surrounding the embryonic implantation sites. However, the roles of AQP7 and the underlying mechanism that regulates AQP7 expression in endometrial decidualisation after implantation are still unclear. This study aimed to investigate the role of the PI3K-Akt pathway in regulating the expression of AQP7 in ESCs and decidualisation. Primary ESCs of pregnant mice were isolated to establish in vitro decidualisation models. PI3K inhibitor LY294002 was added to the decidualisation models, then AQP7 expression, changes in decidualised ESC morphology and expression of decidualisation marker molecules were examined. AQP7 knockdown reduced the proliferation and differentiation of ESCs with in vitro induced decidualisation. Furthermore, when the activity of PI3K was inhibited by LY294002, the expression of AQP7 in decidualised ESCs was decreased and both the proliferation and differentiation of ESCs were significantly reduced. This indicates that AQP7 is a key molecule involved in endometrial decidualisation and the expression of AQP7 is upregulated through activation of the PI3K-Akt pathways, which promotes the proliferation and differentiation of the ESCs, thus affecting occurrence of decidualisation. This study may provide a new biomarker for the diagnosis of infertility and a new drug target for the prevention and treatment of infertility.

Trophectoderm non-coding RNAs reflect the higher metabolic and more invasive properties of young maternal age blastocysts

Increasing female age is accompanied by a corresponding fall in her fertility. This decline is influenced by a variety of factors over an individual’s life course including background genetics, local environment and diet. Studying both coding and non-coding RNAs of the embryo could aid our understanding of the causes and/or effects of the physiological processes accompanying the decline including the differential expression of sub-cellular biomarkers indicative of various diseases. The current study is a post-hoc analysis of the expression of trophectoderm RNA data derived from a previous high throughput study. Its main aim is to determine the characteristics and potential functionalities that characterize long non-coding RNAs. As reported previously, a maternal age-related component is potentially implicated in implantation success. Trophectoderm samples representing the full range of maternal reproductive ages were considered in relation to embryonic implantation potential, trophectoderm transcriptome dynamics and reproductive maternal age. The long non-coding RNA (lncRNA) biomarkers identified here are consistent with the activities of embryo-endometrial crosstalk, developmental competency and implantation and share common characteristics with markers of neoplasia/cancer invasion. Corresponding genes for expressed lncRNAs were more active in the blastocysts of younger women are associated with metabolic pathways including cholesterol biosynthesis and steroidogenesis.

Ultrasound Molecular Imaging of Neovascularization for Evaluation of Endometrial Receptivity Using Magnetic iRGD-Modified Lipid-Polymer Hybrid Microbubbles.

Angiogenesis plays an important role in endometrial receptivity, determining the response of the endometrium to the blastocyst at the early stage of embryo implantation. During the application of assisted reproduction technologies, it is very important to evaluate the status of uterine angiogenesis before deciding on embryo implantation. Targeted microbubbles (MBs)-based ultrasound molecular imaging (UMI) can noninvasively detect the expression status of biomarkers at the molecular level, thereby being a potential diagnosis strategy for various diseases and their therapeutic evaluation. The iRGD-lipopeptide (DSPE-PEG2000-iRGD) conjugate was prepared with iRGD peptides and DSPE-PEG2000-maleimide through the Michael-type addition reaction. Then, the magnetic iRGD-modified lipid-polymer hybrid MBs (Mag-iLPMs) were prepared with the double-emulsification-solvent-evaporation method. Magnetic targeting of Mag-iLPMs was confirmed under the microscope, followed by a rectangular magnet. Next, the in vitro targeted binding of MBs to murine brain-derived endothelial cells.3 (bEnd.3) and human umbilical vein endothelial cells (HUVEC) were evaluated. The ratio of MBs binding to bEnd.3 and HUVEC at the same field was also compared. For in vivo studies, bolus injections of targeted or control MBs were randomly administrated to non-pregnant or pregnant rats on day 5. Then, the uteri were imaged using a VisualSonics Vevo 2100 ultrasound system (Fujifilm VisualSonics Inc., Ontario, Canada) equipped with a high-frequency transducer. Ultrasonic imaging signals were acquired from Mag-iLPMs, and compared with Mag-LPMs, iLPMs, and LPMs. The Mag-iLPMs showed excellent performance in ultrasound contrast imaging and binding affinity to target cells. Using the magnetic field, 10.5- and 12.47-fold higher binding efficiency to bEnd.3 and HUVEC were achieved compared to non-magnetic iLPMs, respectively. Significantly enhanced UMI signals were also observed in the uteri of rats intravenously injected pregnant rats (6.58-fold higher than rats injected with iLPMs). We provided a powerful ultrasonic molecular functional imaging tool for uterine angiogenesis evaluation before embryonic implantation.