Embryonic Development Research Articles

Understanding the effect of acupuncture on nausea and vomiting during pregnancy from a metabolic perspective: study protocol for a single-blinded randomized controlled trial

BackgroundAcupuncture is an effective complementary therapy for nausea and vomiting during pregnancy (NVP). Nevertheless, the utilization of acupuncture for NVP has been minimally explored in current scholarly research, with a paucity of systematic randomized clinical trials (RCTs) in it. We aim to evaluate the effects of acupuncture on NVP after assisted reproductive techniques (ART) and explore the metabolism-related mechanism of the efficacy.MethodsThis single-blind, randomized, controlled trial will randomize 68 patients with NVP after ART to a traditional acupuncture (tACP) or a sham acupuncture (sACP) group. The tACP group will receive tACP thrice a week for 2 weeks with a day interval between sessions, while the sACP group will undergo the same number of nonpenetrative acupuncture at non-acupoints for the same period. Pregnancy-specific quantification of emesis will be used to evaluate symptom severity. Routine blood and urine tests, liver and kidney function tests, human chorionic gonadotropin, nuchal translucency thickness, and embryonic development measured using ultrasound will be used to evaluate safety during pregnancy. Non-targeted metabolomic analysis will be performed to explore the association between metabolic changes and clinical symptoms.DiscussionThis study will elucidate the effects and safety of acupuncture in treating NVP in women undergoing ART. The results of this study will contribute to optimizing acupuncture therapies by combining the body and auricular points and exploring the underlying therapeutic mechanism using a metabolomics approach.Trial registrationChinese Clinical Trial Registry, ChiCTR2300075259.

Prenatal and progressive coenzyme Q10 administration to mitigate muscle dysfunction in mitochondrial disease.

ADCK genesencode aarF domain-containing mitochondrial kinases involved in coenzyme Q (CoQ) biosynthesis and regulation. Haploinsufficiency of ADCK2 in humans leads to adult-onset physical incapacity with reduced mitochondrial CoQ levels in skeletal muscle, resulting in mitochondrial myopathy and alterations in fatty acid β-oxidation. The sole current treatment for CoQ deficiencies is oral administration of CoQ10, which causes only partial recovery with postnatal treatment, underscoring the importance of early diagnosis for successful intervention. We used Adck2 heterozygous mice to examine the influence of this gene on muscle structure, function and regeneration throughout development, growth and ageing. This investigation involved techniques including immunohistochemistry, analysis of CoQ levels, mitochondrial respiratory content, muscle transcriptome analysis and functional tests. We demonstrated that Adck2 heterozygous mice exhibit defects from embryonic development, particularly in skeletal muscle (1102 genes deregulated). Adck2 heterozygous embryos were 7% smaller in size and displayed signs of delayed development. Prenatal administration of CoQ10 could mitigate these embryonic defects. Heterozygous Adck2 mice also showed a decrease in myogenic cell differentiation, with more severe consequences in 'aged' mice (41.63% smaller) (P<0.01). Consequently, heterozygous Adck2 mice displayed accelerated muscle wasting associated with ageing in muscle structure (P<0.05), muscle function (less grip strength capacity) (P<0.001) and muscle mitochondrial respiration (P<0.001). Furthermore, progressive CoQ10 administration conferred protective effects on mitochondrial function (P<0.0001) and skeletal muscle (P<0.05). Our work uncovered novel aspects of CoQ deficiencies, revealing defects during embryonic development in mammals for the first time. Additionally, we identified the gradual establishment and progression of the deleterious Adck2 mouse phenotype. Importantly, CoQ10 supplementation demonstrated a protective effect when initiated during development.

Retraction Note: Zebrafish Noxa promotes mitosis in early embryonic development and regulates apoptosis in subsequent embryogenesis

Retraction Note: Zebrafish Noxa promotes mitosis in early embryonic development and regulates apoptosis in subsequent embryogenesis

Biochemical analysis and fertilization success of Cirrhinus mrigala ova during induced spawning

Fish egg quality is very crucial in aquaculture sector for production of healthy seed. Egg yolk is an energy reservoir for growth and development of embryo. This study evaluated the biochemical composition and quality of Cirrhinus mrigala eggs at three different hatchery sites of Pakistan, (Site 1= Fish Seed Hatchery, District Pakpattan; Site 2= Sidhuwan Hatchery, Head Balloki, District Kasur; Site 3= Chenab Fish Hatchery, Rangpur, District Muzaffargarh) during induced breeding. For this, a total of 36 (18 males and 18 females) fish brooders, 12 (06 males and 06 females) from each site were utilized. Fatty acids analysis revealed significant differences among three different sites. Saturated fatty acids i.e., palmitic acid (C16:0) and stearic acids (C18:0) were higher at site 2 compared to the others sites. Monounsaturated fatty acids i.e., Oleic acid (C18:1) and polyunsaturated fatty acids i.e., Docosahexaenoic acid (DHA) (C22:6) exhibited considerably greater values at site 2 than those of other two sites. Egg mineral contents unveiled remarkable differences, particularly at site 2 indicating significantly higher mineral contents except copper (Cu) in comparison to the other sites. Significant variation exists in fertilization and hatching rates during induced spawning, with the highest values recorded at site 2. It is concluded that biochemical composition of egg especially fatty acid profile and mineral content greatly influences the embryonic development and hatching success of farm reared Cirrhinus mrigala.

Exploring Mechanical Forces Shaping Self-Organization and Morphogenesis During Early Embryo Development.

Embryonic development is a dynamic process orchestrated by a delicate interplay of biochemical and biophysical factors. While the role of genetics and biochemistry in embryogenesis has been extensively studied, recent research has highlighted the significance of mechanical regulation in shaping and guiding this intricate process. Here, we provide an overview of the current understanding of the mechanical regulation of embryo development. We explore how mechanical forces generated by cells and tissues play a crucial role in driving the development of different stages. We examine key morphogenetic processes such as compaction, blastocyst formation, implantation, and egg cylinder formation, and discuss the mechanical mechanisms and cues involved. By synthesizing the current body of literature, we highlight the emerging concepts and open questions in the field of mechanical regulation. We aim to provide an overview of the field, inspiring future investigations and fostering a deeper understanding of the mechanical aspects of embryo development.

Nitrogen-doped carbon quantum dot regulates cell proliferation and differentiation by endoplasmic reticulum stress

ABSTRACT Quantum dots have diverse biomedical applications, from constructing biological infrastructures like medical imaging to advancing pharmaceutical research. However, concerns about human health arise due to the toxic potential of quantum dots based on heavy metals. Therefore, research on quantum dots has predominantly focused on oxidative stress, cell death, and other broader bodily toxicities. This study investigated the toxicity and cellular responses of mouse embryonic stem cells (mESCs) and mouse adult stem cells (mASCs) to nitrogen-doped carbon quantum dots (NCQDs) made of non-metallic materials. Cells were exposed to NCQDs, and we utilized a fluorescent ubiquitination-based cell system to verify whether NCQDs induce cytotoxicity. Furthermore, we validated the differentiation-inducing impact of NCQDs by utilizing embryonic stem cells equipped with the Oct4 enhancer-GFP reporter system. By analyzing gene expression including Crebzf, Chop, and ATF6, we also observed that NCQDs robustly elicited endoplasmic reticulum (ER) stress. We confirmed that NCQDs induced cytotoxicity and abnormal differentiation. Interestingly, we also confirmed that low concentrations of NCQDs stimulated cell proliferation in both mESCs and mASCs. In conclusion, NCQDs modulate cell death, proliferation, and differentiation in a concentration-dependent manner. Indiscriminate biological applications of NCQDs have the potential to cause cancer development by affecting normal cell division or to fail to induce normal differentiation by affecting embryonic development during pregnancy. Therefore, we propose that future biomedical applications of NCQDs necessitate comprehensive and diverse biological studies.

Protein Modifications During Early Embryo Development.

Infertility is a global reproductive health burden. Assisted reproductive technologies (ARTs) have been widely used to help patients become pregnant. Few embryos develop to the blastocyst stage with ARTs, leading to relatively low live birth rates. Protein modifications play crucial roles in nearly every aspect of cell biology, including reproductive processes. The aim of this study was to explore the characteristics of protein modifications during embryonic development. Proteomic data from humans and mice were acquired from the integrated proteome resources (iProX) of ProteomeXchange (PXD024267) and a tandem mass tag (TMT)-mass spectrometry dataset. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were applied for functional annotation. Protein-protein interactions (PPIs) of the modification-related genes were revealed by the STRING database. Modified proteins during mouse embryogenesis were visualized through heatmaps of hierarchically clustering using k-means. We identified modification-related proteins in human embryo development and characterized them through heatmaps, GO analysis, KEGG analysis, and PPI network analysis. We found that the 4-cell stage to the 8-cell stage might be the demarcation period for modification-related protein expression patterns during embryo development. Using quantitative mass spectrometry, we elucidated the methylation, acetylation, and ubiquitination events that occur during mouse embryogenesis to validate our findings in human embryonic development to some extent. The results of our study suggest that the posttranslational modifications (PTMs) of human preimplantation embryos might exhibit the same trends as those in mice to exert synergistic and fine-tuned regulatory effects during embryonic development.

Comparative analysis of embryonic development and growth performance among two mud crab species and their hybrids

In modern aquaculture, inter-specific hybridization enhances traits, but its effects on embryonic development and growth in mud crabs (genus Scylla) remain underexplored. This study investigated the embryonic development and growth performance of Scylla paramamosain, Scylla serrata, and their hybrid offspring from reciprocal crosses. We found that S. paramamosain and the hybrid (S. paramamosain ♀ × S. serrata ♂) produced smaller eggs with shorter development periods than S. serrata and the hybrid (S. serrata ♀ × S. paramamosain ♂). Both hybrids exhibited higher growth rates in the crablet stage than their purebred parents. Notably, the hybrid (S. serrata ♀ × S. paramamosain ♂) demonstrated significant heterosis in body weight and height, while the hybrid (S. paramamosain ♀ × S. serrata ♂) showed single-parent heterosis. While the shape of the frontal lobe spines in hybrids depended on the male parent. Development durations varied, with the longest observed in the hybrid (S. paramamosain ♂ × S. serrata ♀). Discriminant analysis of growth traits showed high accuracy across developmental stages, supporting the potential of inter-specific hybridization for enhancing mud crab aquaculture. This research provides valuable insights and germplasm resources for the aquaculture industry.

Exploring Novel Strategies to Alleviate Symptoms of β-Globinopathies: Examining the Potential Role of Embryonic ε-globin Induction

β-thalassemia and sickle cell disease are among the most prevalent genetic blood disorders globally. These conditions arise from mutations in the β-globin gene, leading to defective hemoglobin production and resulting in anemia. Current treatments include γ-globin inducers (e.g., Hydroxyurea), blood transfusions, iron chelation therapy, and bone marrow transplantation. Recently approved disease-modifying agents and promising gene therapies offer hope, yet their broad application is constrained by scalability challenges. Traditionally, research and development for β-globinopathies have focused on γ-globin induction. However, the ε-globin variant, which is active during early embryonic development and subsequently silenced prenatally, was once considered non-inducible by postnatal pharmacological means. Recent studies indicate that, akin to γ-globin, enhancing ε-globin expression could compensate for impaired β-globin synthesis, potentially ameliorating the clinical manifestations of β-globinopathies. This review critically examines the viability of ε-globin induction as a therapeutic strategy for β-thalassemia and sickle cell diseases. It also delves into the burgeoning research on the mechanisms governing ε-globin silencing and its pharmacological reactivation. We conclude with a discussion of prospective research directions and drug development initiatives aimed at exploiting ε-globin's therapeutic promise.

Brain Derived Neurotrophic Factor Stimulates Hypothalamic and Gonadal Reproductive Hormones and Oocyte Maturation in Zebrafish.

Brain-derived neurotrophic factor (BDNF) is a peptide widely known for its role in neurogenesis and synaptic plasticity. Its expression in non-neuronal tissues has been reported. In mammals, it is involved in ovarian development, follicle growth, oocyte maturation, and early embryonic development. In zebrafish, it was demonstrated that BDNF increases food intake and regulates metabolism. Reproduction and metabolism are tightly linked. We hypothesized that BDNF modulates reproductive hormones and reproductive functions in zebrafish. This study aimed to determine BDNF expression in the zebrafish reproductive axis and whether it modulates the reproductive endocrine milieu and oocyte biology in zebrafish. Our results show that bdnf and its receptor trkb, and BDNF-like immunoreactivity are detected in zebrafish gonads and liver cells. This suggests BDNF local production and possible actions within the gonads and liver. Intraperitoneal administration of 1, 10, or 100 ng/g bodyweight BDNF significantly (ANOVA, p<0.05) increased sgnrh/cgnrh-II, kiss1, and cyp19a1b mRNAs in the zebrafish brain; steroidogenic enzymes (star and cyp19a1a) and key receptors in the zebrafish gonads. In vitro incubation of zebrafish liver cells with BDNF significantly (ANOVA, p<0.05) increased estrogen receptor mRNAs and vitellogenin concentrations (ELISA) in the cells. BDNF (100 ng/mL) induced (ANOVA, p<0.05) oocyte maturation in vitro at 24 hours post-incubation and significantly upregulated cumulus-expansion related genes (ANOVA, p<0.05). Overall, our findings indicate a stimulatory role for BDNF in the reproductive axis of zebrafish. This provides impetus for future research on its mechanism of action and potential practical applications to enhance reproduction in aquaculture.

Low microbial abundance and community diversity in the egg capsule of the oviparous cloudy catshark (Scyliorhinus torazame) during oviposition.

Vertebrate embryos are protected from bacterial infection by various maternally derived factors, yet little is known about the defence mechanisms in elasmobranchs. This study aimed to characterize the intracapsular environment of freshly laid eggs of the oviparous catshark (Scyliorhinus torazame) by investigating the microbial abundance and microbiota to understand its potential contribution to embryonic defence. The egg capsule of oviparous elasmobranchs is tightly sealed until pre-hatching, after which seawater flows into the capsule, exposing the embryos to the surrounding seawater. We found that early embryos were highly vulnerable to environmental pathogens, suggesting that the embryos are somehow protected from infection before pre-hatching. Indeed, the intracapsular environment of freshly laid eggs exhibited significantly low bacterial density, maintained until pre-hatching. Furthermore, the microbiome inside eggs just after oviposition differed markedly from those of rearing seawater and adult oviducal gland epithelia; these eggs were predominantly populated by an unidentified genus of Sphingomonadaceae. Overall, this study provides compelling evidence that early embryos of oviparous cloudy catshark are incubated in a clean intracapsular environment that potentially plays a significant role in embryonic development in oviparous elasmobranchs.

EDC-3 and EDC-4 regulate embryonic mRNA clearance and biomolecular condensate specialization

Animal development is dictated by the selective and timely decay of mRNAs in developmental transitions, but the impact of mRNA decapping scaffold proteins in development is unclear. This study unveils the roles and interactions of the DCAP-2 decapping scaffolds EDC-3 and EDC-4 in the embryonic development of C.elegans. EDC-3 facilitates the timely removal of specific embryonic mRNAs, including cgh-1, car-1, and ifet-1 by reducing their expression and preventing excessive accumulation of DCAP-2 condensates in somatic cells. We further uncover a role for EDC-3 in defining the boundaries between P bodies, germ granules, and stress granules. Finally, we show that EDC-4 counteracts EDC-3 and engenders the assembly of DCAP-2 with the GID (CTLH) complex, a ubiquitin ligase involved in maternal-to-zygotic transition (MZT). Our findings support a model where multiple RNA decay mechanisms temporally clear maternal and zygotic mRNAs throughout embryonic development.

Unveil the toxicity induced on early life stages of zebrafish (Danio rerio) exposed to 3,4-methylenedioxymethamphetamine (MDMA) and its enantiomers

The increased detection of the recreational drug 3,4-methylenedioxymethamphetamine (MDMA) in aquatic ecosystems, has raised concern worldwide about its possible negative impacts on wildlife. MDMA is produced as racemate but its enantioselective effects on non-target organisms are poorly understood. Therefore, this study aimed to provide a comprehensive study of the toxicity of MDMA and its enantiomers in the early life stages of zebrafish (Danio rerio). Zebrafish embryos (≈3 h post fertilization) were exposed to different concentrations (0.02, 0.2, 2, 20, and 200 μg/L) of (R,S)-MDMA and both pure enantiomers.Both enantiomers induced effects on embryonic development, DNA integrity, and behaviour and enantioselective effects were noted. (S)-MDMA exhibits higher toxic effects on embryonic development level with increased mortality and severity of teratogenic effects, and behavioural abnormalities in acoustic startle-habituation response. (R)-MDMA affected general activity and avoidance behaviour, showing greater inhibitory effects on behavioural activity. Additionally, (R,S)-MDMA induced higher genotoxic effects than the two isolated enantiomers. These results are of concern at populational levels since effects on mortality, development, and behaviour were observed even at environmentally relevant concentrations, which can cause a reduction of larval viability and in the number of individuals in each generation, and an increase in the risk of predation of the organisms. Yet, the bioaccumulation studies showed that MDMA is not accumulated in zebrafish.Therefore, this work demonstrated for the first time the occurrence of MDMA enantiotoxicity in the early life stages of zebrafish, which should be considered in further environmental risk assessments involving fish species or other non-target aquatic organisms.

Computing whole embryo strain maps during gastrulation

Gastrulation is a critical process during embryonic development that transforms a single-layered blastula into a multilayered embryo with distinct germ layers, which eventually give rise to all the tissues and organs of the organism. Studies across species have uncovered the mechanisms underlying the building blocks of gastrulation movements, such as localized in-plane and out-of-plane epithelial deformations. The next challenge is to understand dynamics on the scale of the embryo: this requires quantifying strain tensors, which rigorously describe the differences between the deformed configurations taken on by local clusters of cells at time instants of observation and their reference configuration at an initial time. We present a systematic strategy for computing such tensors from the local dynamics of cell clusters, which are chosen across the embryo from several regions whose morphogenetic fate is central to viable gastrulation. As an application of our approach, we demonstrate a strategy of identifying distinct Drosophila morphological domains using strain tensors.

Expression of Wnt signaling proteins in rare congenital bladder disorders

Introduction and AimsCongenital bladder anomalies are rare and are a leading cause of end stage renal failure in children. The Wnt signaling pathway, important during embryonic development, has been implicated in the pathogenesis of these conditions through regulation of gene expression, including essential transcription factors. We investigated the expression of four Wnt transcriptional targets, namely, Pygopus 1 (Pygo1), FRA1, TCF7L1 and Connexin 43 (Cx43) in three rare congenital bladder disorders: bladder exstrophy (BE), neurogenic bladder (NGB) and posterior urethral valves (PUV). MethodsBladder tissue samples were collected from patients at the Great Ormond Street Hospital for Sick Children, London, UK, with control (normally-functioning bladder, N=9), BE (N=15), NGB (N=6) and PUV (N=5). Histological analysis was performed using the van Gieson stain to differentiate smooth muscle (SM) and connective tissue (CT) compartments. An unbiased, automated, semi-quantitative immunofluorescence analysis was performed to measure the labelling intensity of four Wnt-related proteins in tissue from these four groups. Results and DiscussionThere was a significant (p<0.05) increase in the expression of Pygo1 in the smooth muscle of all anomalies examined and also in the connective tissue in PUV compared to control. Cx43 also showed overexpression in the smooth muscle across all conditions; however, there was a reduced expression in NGB and an increase in PUV in connective tissue. TCF7L1 showed a significant decrease in both tissue compartments for NGB, whereas FRA1 expression remained unchanged across all anomalies. We also measured colocalization of Wnt-related proteins. TCF7L1 exhibited increased colocalization with Pygo1 and FRA1 in exstrophy compared to control. These results suggest a complex dysregulation of the Wnt pathway in congenital bladder disorders. ConclusionWnt signaling-related proteins show dysregulation in congenital bladder disorders compared to control tissue. Understanding these mechanisms should help towards non-invasive early diagnosis, drug target discovery and development of treatment strategies for these conditions.Summary tableSummary of Wnt-related protein expression and colocalization in bladder exstrophy (BE), neurogenic bladder (NGB) and posterior urethral valves (PUV) disorder compared to normally-functioning control bladder. The table shows the levels of expression as significantly up (↑) down (↓) or no change (-) of Pygopus 1 (Pygo1), Connexin 43 (Cx43), FRA1 and TCF7L1 expression in smooth muscle and connective tissue compartments in BE, NGB and PUV relative to control bladder tissue samples. The table also shows colocalization of six protein pairs of Pygo1 and FRA1; Pygo1 and Cx43; Pygo1 and TCF7L1; FRA1 and Cx43; FRA1 and TCF7L1 and Cx43 and TCF7L1 in bladder disorders compared to control.Summary tableProtein Expression (Smooth muscle)Pygo1Cx43FRA1TCF7L1BE↑↑--NGB↑↑-↓PUV↑↑--Protein Expression (Connective tissue)BE----NGB-↓-↓PUV↑↑--ColocalizationPygo1/FRA1Pygo1/Cx43Pygo1/TCF7L1FRA1/Cx43FRA1/TCF7L1Cx43/TCF7L1Exstrophy--↑-↑-NGB------PUV------

Conditional heterozygous loss of kit receptor tyrosine kinase in neural crest cell lineage is associated with midline cleft lip and bifid nose deformity

ObjectivesThe receptor tyrosine kinase Kit is expressed in cells derived from the trunk neural crest (NC), such as melanocytes; however, its role in cranial NC cell development is not fully understood. MethodsWe investigated the effects of the heterozygous loss of Kit in NC cells during embryonic development by mating Kit2lox/+ mice with Wnt1-Cre mice to produce Wnt1-Cre; Kit2lox/+ embryos. In addition, Wnt1-Cre mice were mated with Rosa26R-yellow fluorescent protein (YFP) mice to visualize the tissue regions expressing Cre recombinase. Histological studies of the craniofacial regions of these mice were performed using samples from embryonic day (E) 12.5 and postnatal day (P) 1. Cellular apoptosis and proliferation were both analyzed through the immunostaining of tissue sections collected on E13.5 and E14.5 using anti-cleaved caspase 3 (CC3) to detect apoptosis and anti-Ki67 to detect proliferation. Cells from YFP-positive tissue regions of the facial areas of Wnt1-Cre; Kit+/+; Rosa26R–YFP embryos and Wnt1-Cre; Kit2lox/+; Rosa26R–YFP embryos collected on E12.5 and E15.5 were cultured and evaluated for cell proliferation. ResultsCompared with control littermates, Wnt1-Cre; Kit2lox/+ embryos exhibited midline cleft lip and bifid nose deformities. Substantial early (P1) postnatal lethality was observed in Wnt1-Cre; Kit2lox/+ mice, with none surviving to 3 weeks of age. YFP-positive cells from the maxillary regions of Wnt1-Cre; Kit2lox/+; Rosa26R–YFP embryos exhibited defective cell growth and self-renewal in vitro. ConclusionConditional heterozygous loss of Kit in Wnt1-Cre; Kit2lox/+ embryos is associated with craniofacial dysplasia and exhibit defective NC development in vitro and in vivo.

Whole mount preparation and analysis of rabbit mammary gland

The mammary gland undergoes dynamic structural and compositional changes throughout life, influenced significantly by hormonal fluctuations and environmental factors. From embryonic development through menopause, this tissue adapts to accommodate phases such as postnatal expansion, pregnancy-induced lactation, and post-weaning involution. Hormones, growth factors, cytokines, and exogenous factors regulate these innate processes, affecting mammary epithelial cell proliferation and sensitivity, particularly in terminal end buds (TEB) and lobules, which are highly susceptible to endocrine disruption. Rodent models have provided invaluable insights into mammary gland biology, yet differences exist compared to human development, prompting the exploration of alternative models like rabbits. Additionally, there is momentum to move away from the use of nonhuman primates in safety assessments, increasing the need for evaluation tools for all tissues in the rabbit model. Rabbit mammary glands exhibit similarities to humans, making them promising for studying breast biology and pathology. However, protocols for whole-mount analysis of rabbit mammary glands are lacking due to the technical challenges of working with thicker tissue than rodent mammary glands. Here, we developed a methodology modified from rodent studies for preparing and analyzing rabbit mammary gland whole mounts, which is essential for advancing research in mammary gland biology and understanding the effects of hormonal and toxicant-induced disruption of mammary gland growth and function.

Uncovering the association between embryo development and early pregnancy loss using artificial intelligence annotated kinetic events

Research questionCan artificial intelligence powered annotation of numerous biological events help uncover an association between embryonic kinetics and early pregnancy loss? DesignA multicentric retrospective analysis was conducted on 37717 embryos (7028 egg retrievals, 13 centers in France and Spain, three different time-lapse systems). The videos of developing embryos were analyzed with artificial intelligence to detect developmental events, including t2, t5, t8 (cleavage from 2 cells to 8 cells), tSB (start blastulation) and tB (blastocyst). The association between embryo kinetics and transfer outcome (lack of early pregnancy, early pregnancy loss and clinical pregnancy) was investigated using univariate and multivariate logistic regressions. ResultsAn optimal kinetics for clinical pregnancy was identified as being statistically different from that of embryos which led to an early pregnancy loss (p<0.01). As much as overall faster developing embryos (tB-t2=77.18±8.33 hours) were the most likely to lead to an early pregnancy, it appeared that beyond early pregnancy, embryos that showed a deceleration during cleavage (tsB-t5=46.05±13.60 hours) followed by a subsequent acceleration during blastulation (tB-tSB=9.81±5.04 hours) were the most likely to lead to clinical pregnancy. Conversely, embryos that presented the fastest cleavage and longest blastulation kinetics were the most likely to lead to an early pregnancy loss. ConclusionsAnalyzing numerous kinetic combinations annotated with artificial intelligence reveals patterns which can help distinguish embryos that are competent enough for early pregnancy but not for clinical pregnancy. Detecting such subtle kinetic differences could add transparency to algorithms by pinpointing which phases of an embryo development might be predictive of a pregnancy loss.

A new hormonal protocol supports early development of invitro-produced embryos after transfer to anoestrus mares.

The present study aimed to evaluate whether primed anoestrus mares are suitable recipients for embryos produced by intracytoplasmic sperm injection (ICSI). Anoestrus was confirmed in four mares and daily doses of oestradiol benzoate (6 mg in total) over 5 days were administered; after 3 days of rest, oral altrenogest was administered at 0.088 mg/kg and embryos (1 to 5 embryos per mare; 15 in total) were transferred 3.5 days after progesterone onset. Uterine lavage was conducted 48 h after transfer. The results revealed an 80% embryo recovery rate, and among the retrieved embryos, 67% showed evident intrauterine development. Hence, ICSI-derived embryos can be successfully transferred to primed anoestrus mares, but more studies are required to ensure further embryo development and foaling.

Ovarian support cell in vitro maturation (OSC-IVM) results in healthy murine live births with no evidence of reprotoxicology in a multigenerational study

Research questionDoes application of human stem cell-derived ovarian support cells (OSCs) for in vitro maturation (IVM) have a safe reproductive toxicity profile? DesignA total of 70 B6/CBA 6–8-week-old stimulated female mice were used to collect immature oocytes (n=2,025) at the germinal vesicle stage. Oocytes were retrieved denuded (n=930) or with cumulus cells (COCs, n=1,095). These were randomly assigned to traditional IVM media (MediCult-IVMTM, Origio) or the same IVM media (MediCult-IVMTM, Origio) further supplemented with human OSCs (FertiloTM, Gameto Inc.). Metaphase II (M2) oocytes were inseminated by intracytoplasmic sperm injection and cultured to blastocyst stage. Vitrified blastocysts were transferred to pseudopregnant females, and F1 offspring were evaluated for various health and developmental metrics. Select F1 mice were outbred to produce the F2 generation, assessing live births, sex ratio, morphology, behavior, and health. ResultsIn mice OSC-IVM generally led to a decreased maturation rate compared to Traditional-IVM (p=0.0101). Subsequent embryo culture yielded similar cleavage rates between the four groups (p=0.7547). However, there was a significant distinction in the blastocyst formation with OSC-IVM showing higher blastocyst formation rates for both denuded oocytes (p=0.0408) and COCs (p=0.0063). Live births, sex ratio, development, and behavior in F1 and F2 generations revealed no adverse effects or significant differences, confirming the absence of long-term toxicity. ConclusionsThis study demonstrates the safety of the OSC-IVM system, with no adverse effects on embryo development or offspring health and fertility across generations.