Duration of dry and humidified incubation of single-step embryo culture medium and oxygen tension during sham culture do not alter metabolomics signature [version 1; peer review: 1 approved
Ontology highlight
ABSTRACT:
SUBMITTER: Cheredath A
PROVIDER: S-EPMC9214662 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:BackgroundWhole embryo culture is a valuable research method in mammalian developmental biology and birth defects research, enabling longitudinal studies of explanted organogenesis-stage rodent embryos. Rat serum is the primary culture medium, and can sustain growth and development over limited periods as in utero. However, the cost, labor, and time to produce culture serum are factors limiting the uptake of the methodology. The goal of replacing or at least reducing rat serum usage in culture would be in accordance with the principles of "replacement, reduction, and refinement" of animals in research (the 3Rs).MethodsWe performed cultures of mouse embryos for 24 hr from embryonic day 8.5 in serum-free media or in rat serum diluted with defined media, compared with 100% rat serum. Developmental parameters scored after culture included yolk sac circulation, dorsal axial length, somite number, protein content, and completion of cranial neural tube closure.ResultsA literature review revealed use of both serum-free and diluted rat serum-based media in whole embryo culture studies, but with almost no formal comparisons of culture success against 100% rat serum. Two serum-free media were tested, but neither could sustain development as in 100% rat serum. Dilution of rat serum 1:1 with Glasgow Minimum Essential Medium plus defined supplements supported growth and development as well as whole rat serum, whereas other diluent media yielded substandard outcomes.ConclusionRat serum usage cannot be avoided, to achieve high quality mouse embryo cultures, but rat usage can be reduced using medium containing diluted serum.
| S-EPMC6778057 | biostudies-literature
Project description:BackgroundIn this randomized, double-blind, parallel-group trial, we aimed to explore the effectiveness of trigger point dry needling in patients with chronic tension-type headache in reducing headache frequency, intensity and duration, and improvement of health-related quality of life.MethodsThe 168 patients in 2 neurology clinics with chronic tension-type headache. The participants were randomly assigned to one of two treatment groups for dry needling or sham dry needling, delivered in 3 sessions a week for 2 weeks. The 160 patients fulfilled the study requirements. The dry needling was applied in active trigger points located in the musculature of the head and the neck. The patients received dry needling using sterile stainless-steel acupuncture needles of 0.25 × 40 mm and 0.25 × 25 mm dimensions. The sham dry needling procedure was applied into the adipose tissue located at any area where an active trigger point was absent. The primary outcome measurement was the headache intensity. Secondary outcomes were frequency and duration of headache, and quality of life, assessed by the Short Form-36. All outcomes were measured at baseline, at the end of 2-week, and 1-month follow-up period.ResultsIn the dry needling group, intensity, frequency and duration of headache, and the scores of Short Form-36 subscales were significantly improved after treatment (P < .05). In the dry needling group, all the effect sizes for headache variables were large.ConclusionsThe results of this clinical trial suggest that trigger point dry needling in patients with chronic tension-type headache is effective and safe in reducing headache intensity, frequency and duration, and increasing health-related quality of life.Trial registrationClinical Trials NCT03500861.
| S-EPMC6408118 | biostudies-literature
Project description:BackgroundMetabolites in spent embryo culture medium correlate with the embryo's viability. However, there is no widely accepted method using metabolite dada to predict successful implantation. We sought to combine metabolomic profiling of spent embryo culture medium and clinical variables to create an implantation prediction model as an adjunct to morphological screening of day 3 embryos.MethodsThis investigation was a prospective, nested case-control study. Forty-two day 3 embryos from 34 patients were transferred, and the spent embryo culture medium was collected. Twenty-two embryos implanted successfully, and the others failed. Metabolites in the medium relevant to implantation were detected and measured by Liquid Chromatography-Mass Spectrometry. Clinical signatures relevant to embryo implantation were subjected to univariate analysis to select candidates for a prediction model. Multivariate logistical regression of the clinical and metabolomic candidates was used to construct a prediction model for embryo implantation potential.ResultsThe levels of 13 metabolites were significantly different between the successful and failed groups, among which five were most relevant and interpretable selected by Least Absolute Shrinkage and Selection Operator regression analysis. None of the clinical variables significantly affected day 3 embryo implantation. The most relevant and interpretable set of metabolites was used to construct a prediction model for day 3 embryo implantation potential with an accuracy of 0.88.ConclusionsDay 3 embryos'implantation potential could be noninvasively predicted by the spent embryo culture medium's metabolites measured by LC-MS. This approach may become a useful adjunct to morphological evaluation of day 3 embryos.
| S-EPMC10249307 | biostudies-literature
Project description:Hypotaurine (HT) is a routine component of porcine embryo culture medium, functioning as an antioxidant, but its requirement may be diminished as most embryo culture systems now use 5% O2 instead of atmospheric (20%) O2 . Our objective was to determine the effects of removing HT from the culture medium on porcine preimplantation embryo development. Embryos cultured in 20% O2 without HT had decreased blastocyst development compared to culture with HT or in 5% O2 with or without HT. Notably, differences in blastocyst development or total cell number were not detected between embryos cultured in 5% O2 with or without HT. After culture in 5% O2 without HT and embryo transfer, healthy fetuses were retrieved from two pregnancies on Day 42, confirming in vivo developmental competence. Transcript abundance of proapoptotic markers was decreased in embryos cultured without HT regardless of oxygen tension; however, assays for apoptosis did not demonstrate differences between groups. Additionally, no differences were observed in the development or apoptosis of somatic cell nuclear transfer-derived embryos cultured in 5% O2 with or without HT. With decreased utility in 5% O2 , removing HT from porcine embryo culture medium would also have economic advantages because it is undoubtedly the most expensive component.
| S-EPMC7496716 | biostudies-literature
Project description:PURPOSE:The aim of this prospective randomized control trial was to evaluate if the use of two different volumes (20-25 vs 40-45 ?l) of media used for embryo transfer affects the clinical outcomes in fresh in vitro fertilization (IVF) cycles. METHODS:In total, 236 patients were randomized in two groups, i.e., "low volume" group (n?=?118) transferring the embryos with 20-25 ?l of medium and "high volume" group (n?=?118) transferring the embryos with 40-45 ?l of medium. The clinical pregnancy, implantation, and ongoing pregnancy rates were compared between the two groups. RESULTS:No statistically significant differences were observed in clinical pregnancy (46.8 vs 54.3%, p?=?0.27), implantation (23.7 vs 27.8%, p?=?0.30), and ongoing pregnancy (33.3 vs 40.0%, p?=?0.31) rates between low and high volume group, respectively. CONCLUSION:Higher volume of culture medium to load the embryo into the catheter during embryo transfer does not influence the clinical outcome in fresh IVF cycles. TRIAL REGISTRATION NUMBER:NCT03350646.
| S-EPMC5949098 | biostudies-literature
Project description:BackgroundGiven implantation failure limits the improvement of in vitro fertilization (IVF) success rates, there is an urgency to identify potential biomarkers for embryo quality and predict the outcomes of IVF-embryo transfer (IVF-ET).MethodsUsing RNA-sequencing, we identified the expression profiles of 16 spent culture medium (SCM) collected from embryos at the cleavage on day 3 (D3 cleavage) and blastocyst stages on day 5 (D5 blastocyst) during IVF cycles. Differentially expressed miRNAs (DEmiRNAs) were then identified, and microRNA (miRNA)-messenger RNA (mRNA) interaction networks were constructed. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) confirmation and validation in the Gene Expression Omnibus (GEO) database were performed.ResultsCompared with the pregnant group, 29 DEmiRNAs were detected in the non-pregnant group at D3 cleavage, and 26 were detected in the non-pregnant group at D5 blastocyst. Among them, a total of six known miRNAs, including hsa-miR-199a-3p>hsa-miR-199b-3p, hsa-miR-199a-5p, hsa-miR-379-5p, hsa-miR-432-5p, hsa-miR-99a-5p, and hsa-miR-483-5p, were identified. The results of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these target genes of DEmiRNAs were associated with various biological processes (BPs). The results of validation in qRT-PCR and the GEO database suggested the reliability of our RNA-sequencing results.ConclusionsIn conclusion, we identified three miRNAs, including hsa-miR-199a-5p, hsa-miR-483-5p, and hsa-miR-432-5p, which may serve as biomarkers for embryo quality during IVF cycles.
| S-EPMC8667117 | biostudies-literature
Project description:The oil overlay micro-drop system is widely used for cultures of mammalian gametes and embryos. We evaluated hereby the effects of two unaltered commercial oils- Sigma mineral oil (S-MO) and Nidoil paraffin oil (N-PO)-on in vitro embryo production (IVP) outcomes using a pig model. The results showed that while either oil apparently did not affect oocyte maturation and fertilization rates, S-MO negatively affected embryo cleavage rates, blastocyst formation rates, and, consequently, total blastocyst efficiency of the system. No differences in the oxidation state were found between the oils or culture media incubated under S-MO or N-PO. Although both oils slightly differed in elemental composition, there were no differences in the concentrations of elements between fresh media and media incubated under oils. By contrast, we demonstrated clear oil-type differences in both the composition of volatile organic compounds (VOC) and the transfer of some of these VOC´s (straight-chain alkanes and pentanal and 1,3-diethyl benzene) to the culture medium, which could have influenced embryonic development.
| S-EPMC5585310 | biostudies-literature
Project description:Mechanotransduction, the process by which cells respond to tension transmitted through various supramolecular linkages, is important for understanding cellular behavior. Tension gauge tethers (TGTs), short fragments of double-stranded DNA that irreversibly break under shear-stretch conditions, have been used in live cell experiments to study mechanotransduction. However, our current understanding of TGTs' mechanical responses is limited, which limits the information that can be gleaned from experimental observations. In this study, we quantified the tension-dependent lifetime of TGTs to better understand their mechanical stability under various physiologically relevant stretching conditions. This work has broad applications for using TGTs as tension threshold and duration sensors and also suggests the need to revisit previous interpretations of experimental observations.
| S-EPMC9973368 | biostudies-literature
Project description:The metabolomic profile of an embryo culture medium can aid in the advanced prediction of embryonic developmental potential and genetic integrity. But it is not known if this technology can be used to determine the in vitro potential of inner cell mass (ICM) in adherence and proliferation. Here, we investigated the developmental potential of mouse 2-cell embryos carrying cisplatin-induced DNA lesions (IDL), beyond blastocyst stage using ICM outgrowth assay. The genetic integrity of ICM cells was determined by comet assay. The metabolic signatures of spent medium were recorded 84 hours post injection of hCG (hpi-hCG), and after 96 hours of extended in vitro culture (Ex 96) by NMR spectroscopy. We observed that blastocysts that lack the ability to adhere in vitro had an increased requirement of pyruvate (p < 0.01), lactate (p < 0.01), and were accompanied by a significant reduction of pyruvate-alanine ratio in the culture medium. We propose that the aforementioned metabolites from 84 hpi-hCG spent medium be further explored using appropriate experimental models, to prove their potential as biomarkers in the prediction of implantation ability of in vitro derived human embryos in clinical settings.
| S-EPMC6242932 | biostudies-literature
Project description:The HLA-G molecule is widely accepted as an important factor for pregnancy success. Its expression has been detected in the extravillous trophoblasts. Soluble HLA-G (sHLA-G) was found in the genital tract, pre-implanted embryos as well as in seminal fluid. In this study, we investigated the concentration of sHLA-G (sHLA-G1 and sHLA-G5) in media from 344 single cultured embryos following in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). The level of sHLA-G (U/ml) was tested with a sandwich enzyme-linked immunosorbent assay (ELISA) kit. We correlated sHLA-G secretion with ovarian stimulation protocols, the type of embryo transfer (fresh or frozen cycle) and the quality of the embryos. The ovarian stimulation protocol affects the secretion of sHLA-G by the embryo. Embryos obtained from the long agonist protocol secreted more sHLA-G than those originating from the short antagonist protocol (p = 0.0001). Embryos whose transfer resulted in a clinical pregnancy and/or live birth secreted more sHLA-G compared to those whose transfer ended without pregnancy. This was particularly observable in embryos following the long ovarian stimulation protocol and from a frozen embryo cycle. In conclusion, sHLA-G secreted by the embryo has an impact on implantation and live birth and could be a developmental potential marker of the embryo. Its concentration depends on the ovarian stimulation protocol used.
| S-EPMC9730522 | biostudies-literature