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Mastenbroek, S.

Publications and source records attributed to Mastenbroek, S..

4 recordsLinked to original sources

In vivo human uterine temperature, pH, and uterine fluid composition analysis

Study questionWhat are the temperature, pH and uterine fluid composition in the human uterus three days following a positive LH test or ovum pick-up? Summary answerThe mean uterine temperature was 36.94{+/-}0.26{degrees}C, the mean uterine pH was 6.76{+/-}0.22, and the concentrations of 37 components in aspirated uterine fluid were successfully determined. What is known alreadyEmbryo culture conditions in the laboratory impact key outcomes of IVF/ICSI treatments, such as the quality of the embryos and the live birth rate after treatment, and child outcomes, such as birth weight. Currently used conditions, including temperature, pH, and culture medium composition, are largely derived from clinical experience and experimental studies using animal models. Limited studies have been performed to determine the natural human preimplantation embryo environment in vivo during the physiologically relevant time of the menstrual cycle. This type of fundamental knowledge is required for evidence-based optimization of the in vitro embryo culture environment and improving IVF/ICSI outcomes. Study design, size, durationIn this cross-sectional study, conducted between April 2015 and March 2016, temperature and pH were measured in the human uterine cavity on the third day following a positive LH test or ovum pick-up, and uterine fluid was simultaneously aspirated for composition analysis. Uterine temperature was measured in fifty eight women, uterine pH was determined in fifty three women, and twenty two samples of aspirated uterine fluid were analysed for the concentrations of thirty-seven components. Participants/materials, setting, methodsThis study involved 61 healthy reproductive-aged women: 53 without ovarian stimulation and 8 who underwent ovarian stimulation. We measured uterine temperature using a probe inserted into the uterine cavity directly, and uterine pH after inserting a probe through the outer sheath of an IVF catheter. Uterine fluid was then aspirated using this outer IVF catheter and a 10 ml syringe, and subsequently analysed with a Cobas 8000 chemistry analyser and ultra-performance liquid chromatography-tandem mass spectrometry. Main results and the role of chanceThe mean uterine temperature on the third day following a positive LH test or ovum pick-up was 36.94 {+/-} 0.26{degrees}C and correlated with the womens core body temperature. The mean pH in the uterine cavity was pH 6.76 {+/-} 0.22, clearly lower than the standard pH used for human preimplantation embryo culture in vitro (pH 7.3 {+/-} 0.1). Concentrations of important energy sources were 0.8 {+/-} 0.02 mM pyruvate, 5.1 {+/-} 1.78 mM glucose and 6.60 {+/-} 1.12 mM lactate. Glutamic acid (1162 {+/-} 183 M), glycine (955 {+/-} 156 M) and alanine (513 {+/-} 82 M) were the most abundant amino acids in uterine fluid. Limitations, reasons for cautionIn absence of a preimplantation embryo, synergistic influences on the uterine environment may be overlooked. Single centre and specific population limitations may hinder broader generalization of the results. Uterine fluid likely contains additional components. Wider implications of the findingsThe in vivo uterine characteristics identified in this study are foundational to develop an in vivo evidence-based culture medium for human embryos. Further research is necessary to evaluate whether such a medium can improve human preimplantation embryo development and quality, thereby increasing cumulative live birth rates and improving child outcomes.

developmental biology↗

Human blastocyst formation in an embryo culture medium mimicking human uterine pH and fluid composition

Study questionTo study the development of human embryos in a culture medium with pH 6.8 and composition based on previous in vivo human uterine measurements. Summary answerThe blastocyst formation rate of donated surplus human preimplantation embryos in embryo culture medium with pH 6.8 and uterine composition was similar to control medium. What is known alreadyIn vitro culture conditions can affect human preimplantation embryo development and fertility treatment success rates. The current IVF culture conditions are derived from using animal models, laboratory experience and limited human in vivo evidence. Therefore, we previously measured human in vivo uterine conditions on the third day following a positive LH test or ovum pick-up. Strikingly, the uterine pH at this time of the menstrual cycle (pH 6.8) appeared lower than the pH used for human preimplantation embryo culture in IVF laboratories worldwide (pH 7.2-7.4). Mimicking in vivo conditions may potentially improve in vitro embryo development, implantation rates, and overall fertility treatment and safety outcomes. Study design, size, durationIn this preclinical pilot study, 404 donated surplus human day 3 and day 4 embryos were cultured in one of four culture media: ULPC medium (uterine low pH and uterine fluid composition), UC medium (standard pH and uterine fluid composition), ULP medium (uterine pH and standard composition), or control medium (G-2 PLUS medium with standard pH and composition). Blastocyst formation rate on day 5 (after one or two days in culture) was the primary outcome to assess embryo development. Participants/materials, setting, methodsEmbryos were randomly allocated to the different culture conditions, stratified by sibling status and maternal age, before thawing. Day 3 and day 4 embryos were separately randomized, ensuring that each culture group contained an equal amount of both day 3 and day 4 embryos. All procedures and culture conditions were similar between groups, except for pH and medium composition. Embryo morphology was assessed immediately after thawing and on developmental day 5 for the primary outcome. Assessors were blinded to medium allocation. Main results and the role of chanceAll four culture media supported human preimplantation embryo development into blastocysts. Blastocyst formation rates on day five were 54/102 (52.9%) in ULPC medium, 44/104 (42.3%) in UC medium, 48/98 (49.0%) in ULP medium and 47/102 (46.1%) in control medium. Limitations, reasons for cautionThis study is limited by the use of donated surplus human day three and day four embryos. The effect of these new in vitro conditions on embryo development during the first three days of human embryo culture is not yet known. Wider implications of the findingsThe outcomes of this study indicate the need for further research on mimicking the in vivo uterine conditions for in vitro embryo culture using the conditions proposed here, including a lower pH (of 6.8) than currently used in IVF laboratories (pH 7.2-7.4).

developmental biology↗

Human ovarian ageing is characterized by oxidative damage and mitochondrial dysfunction

Human ovarian ageing encompasses the age-related decline in female fertility. Oxidative stress and mitochondrial dysfunction in oocytes are suggested as causal, but corroborating evidence is limited. Using immunofluorescence imaging on human ovarian tissue, we found oxidative damage by protein and lipid (per)oxidation at the primordial follicle stage. Additionally, using comprehensive metabolomics and lipidomics, a cohort of 150 human germinal vesicles and metaphase I oocytes and 15 corresponding cumulus cell samples displayed a shift in glutathione to oxiglutathione ratio and depletion of phospholipids. Age-related changes in polar metabolites suggested a decrease in mitochondrial function, as demonstrated by NAD+, purine and pyrimidine depletion, while glycolysis substrates and glutamine accumulated with age. Oocytes of advanced maternal age likely used alternative energy sources like glycolysis and the adenosine salvage pathway, and possibly increased ATP production in cumulus cells. These findings indicate that oocytes of advanced maternal age suffer from oxidative damage and mitochondrial dysfunction. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/525662v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1d4e4f5org.highwire.dtl.DTLVardef@397eborg.highwire.dtl.DTLVardef@1eacf90org.highwire.dtl.DTLVardef@e13471_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

High-quality human preimplantation embryos stimulate endometrial stromal cell migration via secretion of microRNA hsa-miR-320a

Implantation failure is one of the major success limiting factors in human reproduction. Despite, the mechanisms that determine successful human embryo implantation remain largely unknown. We here show that high-quality human preimplantation embryos secrete soluble signaling factors, including micro RNA (miRNA) hsa-miR-320a, that promote migration of human endometrial stromal cells (hESCs). By using miRNA mimics and inhibitors, we demonstrate that hsa-miR-320a alone can stimulate migration of decidualized hESCs, accurately resembling the response typically triggered only by high-quality embryos. Transcriptome analysis further demonstrated that this effect is very likely mediated via altered expression of genes involved in cell adhesion and cytoskeleton organization. In conclusion, by secreting hsa-miR-320a, high-quality human preimplantation embryos directly influence endometrial stromal cells, most likely to prime the endometrium at the implantation site for successful implantation. Together, our results indicate that hsa-miR-320a may be a promising target to further increase success rates in assisted reproduction.

developmental biology↗