Embryo Model Research Articles

Human embryo models made from pluripotent stem cells are not synthetic; they aren't embryos, either.

Embryo models are potentially highly impactful for human health research because their development recapitulates otherwise inaccessible events in a poorly understood area of biology, the first few weeks of human life. Casual reference to these models as "synthetic embryos" is misleading and should be approached with care and deliberation.

Antimelanogenic potential of brewer's spent grain extract through modulation of the MAPK/MITF axis

Beer brewing produces >39 million tons of brewer's spent grain (BSG) per year worldwide. Although BSG is rich in protein, fiber, and phytochemicals, it is mainly used as livestock feed, and some is discarded. In this study, we attempted to assess the antimelanogenesis effects of BSG and identify the functional components. To this end, we investigated the antioxidant and antimelanin synthesis effects of a BSG extract (BSGE). The results showed that BSGE has high DPPH radical-scavenging activity and inhibited α-MSH-induced melanin synthesis in B16F10 cells cultured in both 2D and 3D environments. BSGE also reduced the expression levels of MITF, TRP-1, TRP-2, and tyrosinase in B16F10 cells by regulating the MAPK pathway. We confirmed the skin-whitening effect of BSGE in 3D reconstituted human skin and zebrafish embryo models. HPLC analysis showed that BSGE contains ferulic acid and p-coumaric acid, which are known to have whitening effects. Thus, BSGE can be used in multifunctional cosmetics as a sustainable ingredient with antioxidant and skin-whitening properties.

4G mobile phone radiation alters some immunogenic and vascular gene expressions, and gross and microscopic and biochemical parameters in the chick embryo model.

The risks to human health have grown over the past 10 years due to the excessive use of mobile phones. The study was designed to determine the harmful effects of 4G mobile phone radiation on the expression of immunogenic and vascular genes and gross, microscopic and biochemical alterations in the development of chicken embryos. Sixty individuals in the exposure group were subjected to mobile phones with a specific absorption rate of 1.4W/kg and a frequency of 2100MHz positioned at a distance of 12cm in the incubator for 60min/night for 14 days. The histopathological examination involved hematoxylin and eosin staining, whereas cresyl violet staining was used to evaluate the condition and number of neurons in the brain. The biochemical parameters of amniotic fluid were analysed using the photometry method, and the expression of VEGF-A and immunity genes (AvBD9, IL6) was measured using the real-time PCR (qPCR) technique. Compared to the control, the exposure group's body weight and length significantly decreased (p<0.05). Subcutaneous bleeding was seen in the exposure group. Urea, creatinine, alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase levels were all significantly higher than in the control group (p<0.05). The exposed group showed pathological lesions in the liver and degenerated neurons with lightly stained nuclei in the cerebral cortex. Hyperchromatic neurons were significantly higher in the exposure group (58.8±2.28) compared to the control (6.6±0.44) (p<0.05). 4G exposure reduced lymphocyte count in the caecal tonsil (86.8±5.38) compared to the control (147.2±9.06) (p<0.05). Vascular gene mRNA expression was higher, but immune gene expression was lower in the exposed group. Exposure to mobile phone radiation may result in gross, microscopic and biochemical changes, as well as alterations in gene expression that could hinder embryonic development.

Glycerol monolaurate improves intestinal morphology and antioxidant status by suppressing inflammatory responses and nuclear factor kappa-B signaling in lipopolysaccharide-exposed chicken embryos

Medium-chain fatty acids and their derivatives are natural ingredients that support immunological functions in animals. The effects of glycerol monolaurate (GML) on intestinal innate immunity and associated molecular mechanisms were investigated using a chicken embryo model. Sixty-four Arbor Acres broiler embryos were randomly allocated into four groups. On embryonic day 17.5, the broiler embryos were administered with 9 mg of GML, which was followed by a 12-h incubation period and a 12-h challenge with 32 μg of lipopolysaccharide (LPS). On embryonic day 18.5, the jejunum and ileum were harvested. Results indicated that GML reversed the LPS-induced decline in villus height and upregulated the expression of mucin 2 (P < 0.05). GML decreased LPS-induced malondialdehyde production and boosted antioxidant enzyme activity (P < 0.05). GML alleviated LPS-stimulated intestinal secretion of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) (P < 0.05). GML also normalized LPS-induced changes in the gene expression of Toll-like receptor 4, nuclear factor kappa-B p65 (NF-κB p65), cyclooxygenase-2, NOD-like receptor protein 3, IL-18, zonula occludens 1, and occludin (P < 0.05). GML enhanced as well the expression of AMP-activated protein kinase α1 and claudin 1 (P < 0.05). In conclusion, GML improved intestinal morphology and antioxidant status by alleviating inflammatory responses and modulating NF-κB signaling in LPS-challenged broiler embryos.

Wnt and BMP signalling direct anterior-posterior differentiation in aggregates of mouse embryonic stem cells.

Stem-cell-based embryo models have allowed greater insight into peri-implantation mammalian developmental events that are otherwise difficult to manipulate due to the inaccessibility of the early embryo. The rapid development of this field has resulted in the precise roles of frequently used supplements such as N2, B27 and Chiron in driving stem cell lineage commitment not being clearly defined. Here, we investigate the effects of these supplements on embryoid bodies to better understand their roles in stem cell differentiation. We show that Wnt signalling has a general posteriorising effect on stem cell aggregates and directs differentiation towards the mesoderm, as confirmed through the upregulation of posterior and mesodermal markers. N2 and B27 can mitigate these effects and upregulate the expression of anterior markers. To control the Wnt gradient and the subsequent anterior versus posterior fate, we make use of a BMP4 signalling centre and show that aggregates in these conditions express cephalic markers. These findings indicate that there is an intricate balance between various culture supplements and their ability to guide differentiation in stem cell embryo models.

Embryo model completes gastrulation to neurulation and organogenesis

Embryo model completes gastrulation to neurulation and organogenesis

Complete human day 14 post-implantation embryo models from naive ES cells

The ability to study human post-implantation development remains limited owing to ethical and technical challenges associated with intrauterine development after implantation1. Embryo-like models with spatially organized morphogenesis and structure of all defining embryonic and extra-embryonic tissues of the post-implantation human conceptus (that is, the embryonic disc, the bilaminar disc, the yolk sac, the chorionic sac and the surrounding trophoblast layer) remain lacking1,2. Mouse naive embryonic stem cells have recently been shown to give rise to embryonic and extra-embryonic stem cells capable of self-assembling into post-gastrulation structured stem-cell-based embryo models with spatially organized morphogenesis (called SEMs)3. Here we extend those findings to humans using only genetically unmodified human naive embryonic stem cells (cultured in human enhanced naive stem cell medium conditions)4. Such human fully integrated and complete SEMs recapitulate the organization of nearly all known lineages and compartments of post-implantation human embryos, including the epiblast, the hypoblast, the extra-embryonic mesoderm and the trophoblast layer surrounding the latter compartments. These human complete SEMs demonstrated developmental growth dynamics that resemble key hallmarks of post-implantation stage embryogenesis up to 13–14 days after fertilization (Carnegie stage 6a). These include embryonic disc and bilaminar disc formation, epiblast lumenogenesis, polarized amniogenesis, anterior–posterior symmetry breaking, primordial germ-cell specification, polarized yolk sac with visceral and parietal endoderm formation, extra-embryonic mesoderm expansion that defines a chorionic cavity and a connecting stalk, and a trophoblast-surrounding compartment demonstrating syncytium and lacunae formation. This SEM platform will probably enable the experimental investigation of previously inaccessible windows of human early post implantation up to peri-gastrulation development.

Alternatives to Fish Acute Whole Effluent Toxicity (WET) Testing: Predictability of RTgill-W1 Cells and Fathead Minnow Embryos with Actual Wastewater Samples.

Toxicity assays using fish cells and embryos continue to gain momentum as a more ethical and informative alternative to fish acute toxicity testing. The goal of our study was to test the accuracy of RTgill-W1 cells and the fathead minnow (Pimephales promelas) embryos to predict actual whole effluent toxicity (WET) in the fathead minnow larvae. The three models were compared concurrently using samples of various origins and treatment types. Additionally, the toxicity of reference toxicants (Cd, Cu, NH3-N, 3,4-dichloraniline, and benzalkonium chloride) spiked into a nontoxic wastewater was compared. The toxicity of reference toxicants was tested in isosmotic and hypoosmotic exposure media in RTgill-W1 cells. Of the 28 wastewater samples, 14 induced a toxic response in fish larvae. Embryos predicted 11 of the 14 wastewater samples toxic to the larvae, whereas RTgill-W1 cells predicted the toxicity of all 14 toxic samples to the larvae. In addition, embryos and RTgill-W1 cells predicted toxicity in two and six additional samples, respectively, that were nontoxic to larvae. Exposures in hypoosmotic medium significantly increased sensitivity of RTgill-W1 cells to all reference toxicants, excluding benzalkonium chloride, compared to exposures in isosmotic medium and showed toxicity levels similar to that in larvae. Thus, hypoosmotic exposure medium should be considered for aquatic toxicity testing applications. Overall, both gill cell and embryo models predicted toxicity in the majority of wastewater samples toxic to larvae and demonstrated their applicability for regulatory WET testing.

In Vitro Embryogenesis and Gastrulation Using Stem Cells in Mice and Humans.

During early mammalian embryonic development, fertilized one-cell embryos develop into pre-implantation blastocysts and subsequently establish three germ layers through gastrulation during post-implantation development. In recent years, stem cells have emerged as a powerful tool to study embryogenesis and gastrulation without the need for eggs, allowing for the generation of embryo-like structures known as synthetic embryos or embryoids. These in vitro models closely resemble early embryos in terms of morphology and gene expression and provide a faithful recapitulation of early pre- and post-implantation embryonic development. Synthetic embryos can be generated through a combinatorial culture of three blastocyst-derived stem cell types, such as embryonic stem cells, trophoblast stem cells, and extraembryonic endoderm cells, or totipotent-like stem cells alone. This review provides an overview of the progress and various approaches in studying in vitro embryogenesis and gastrulation in mice and humans using stem cells. Furthermore, recent findings and breakthroughs in synthetic embryos and gastruloids are outlined. Despite ethical considerations, synthetic embryo models hold promise for understanding mammalian (including humans) embryonic development and have potential implications for regenerative medicine and developmental research.

P-160 - Combination of JAKi and HDACi as a potential anti-angiogenic treatment in a cutaneous T-cell lymphoma (CTCL) in vivo chick embryo model

P-160 - Combination of JAKi and HDACi as a potential anti-angiogenic treatment in a cutaneous T-cell lymphoma (CTCL) in vivo chick embryo model

In preprints: opportunities to unravel the earliest stages of human development using stem cell-based embryo models.

In preprints: opportunities to unravel the earliest stages of human development using stem cell-based embryo models.

Characterization and In Vivo Antiangiogenic Activity Evaluation of Morin-Based Cyclodextrin Inclusion Complexes.

Morin (MRN) is a natural compound with antiangiogenic, antioxidant, anti-inflammatory, and anticancer activity. However, it shows a very low water solubility (28 μg/mL) that reduces its oral absorption, making bioavailability low and unpredictable. To improve MRN solubility and positively affect its biological activity, particularly its antiangiogenic activity, in this work, we prepared the inclusion complexes of MNR with sulfobutylether-β-cyclodextrin (SBE-β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD). The inclusion complexes obtained by the freeze-drying method were extensively characterized in solution (phase-solubility studies, UV-Vis titration, and NMR spectroscopy) and in the solid state (TGA, DSC, and WAXD analysis). The complexation significantly increased the water solubility by about 100 times for MRN/HP-β-CD and 115 times for MRN/SBE-β-CD. Furthermore, quantitative dissolution of the complexes was observed within 60 min, whilst 1% of the free drug dissolved in the same experimental time. 1H NMR and UV-Vis titration studies demonstrated both CDs well include the benzoyl moiety of the drug. Additionally, SBE-β-CD could interact with the cinnamoyl moiety of MRN too. The complexes are stable in solution, showing a high value of association constant, that is, 3380 M-1 for MRN/HP-β-CD and 2870 M-1 for MRN/SBE-β-CD. In vivo biological studies on chick embryo chorioallantoic membrane (CAM) and zebrafish embryo models demonstrated the high biocompatibility of the inclusion complexes and the effective increase in antiangiogenic activity of complexed MRN with respect to the free drug.

Photothermally Enhanced Dual Enzyme‐mimic Activity of Gold‐Palladium Hybrid Nanozyme for Cancer Therapy

Comprehensive SummaryBased on characteristics of the tumor microenvironment (TME), including acidity, hypoxia, inflammation and hydrogen peroxide overload, combined with emerging nanotechnologies, designing nanoplatforms with TME specificity/responsiveness for tumor treatment is a promising nanotherapeutic strategy. In this work, a multifunctional gold‐palladium bimetallic cascade nanozyme was constructed for effective photothermal‐enhanced cascade catalyzed synergistic therapy of tumors. The dumbbell‐like Au‐Pd bimetallic nanomaterial (Au NRs‐Pd@HA) was obtained by reducing palladium on gold nanorods with ascorbic acid (AA) and further modified with hyaluronic acid (HA). The introduction of HA brings biocompatibility and targeting properties. The zebrafish embryos model showed that Au NRs‐Pd@HA had good biocompatibility and low biotoxicity. Au NRs‐Pd@HA can induce catalytic conversion of glucose to generate H2O2 efficiently, and subsequently undergo cascade reaction to produce abundant ·OH radicals, exhibiting peroxidase‐like (POD‐like) and glucose oxidase‐like (GOD‐like) capabilities. The generated ·OH was a key factor for tumor ablation. Meanwhile, Au NRs‐Pd@HA exhibits good photothermal performance under 808 nm irradiation, in favor of photothermal therapy (PTT). Especially, the POD‐like and GOD‐like activities were significantly enhanced due to the photothermal effect. The synergistic PTT and photothermal‐enhanced nanozymes with cascade catalytic effect enabled efficient and safe cancer therapy.

Anti-Melanogenic and Antioxidant Activity of Bifidobacterium longum Strain ZJ1 Extracts, Isolated from a Chinese Centenarian.

Melanin produced by melanocytes protects our skin against ultraviolet (UV) radiation-induced cell damage and oxidative stress. Melanin overproduction by hyperactivated melanocytes is the direct cause of skin hyperpigmentary disorders, such as freckles and melasma. Exploring natural whitening agents without the concern of toxicity has been highly desired. In this study, we focused on a Bifidobacterium longum strain, ZJ1, isolated from a Chinese centenarian, and we evaluated the anti-melanogenic activity of the distinctive extracts of ZJ1. Our results demonstrated that whole lysate (WL) and bacterial lysate (BL) of ZJ1 ferments efficiently reduce α-melanocyte-stimulating hormone (α-MSH)-induced melanin production in B16-F10 cells as well as the melanin content in zebrafish embryos. BL and WL downregulate melanogenesis-related gene expression and indirectly inhibit intracellular tyrosinase activity. Furthermore, they both showed antioxidant activity in a menadione-induced zebrafish embryo model. Our results suggest that ZJ1 fermentation lysates have application potential as therapeutic reagents for hyperpigmentary disorders and whitening agents for cosmetics.

Impact of Drug Administration Routes on the In Vivo Efficacy of the Natural Product Sorangicin a Using a Staphylococcus aureus Infection Model in Zebrafish Embryos.

Staphylococcus aureus causes a wide range of infections, and it is one of the leading pathogens responsible for deaths associated with antimicrobial resistance, the rapid spread of which among S. aureus urges the discovery of new antibiotics. The evaluation of in vivo efficacy of novel drug candidates is usually performed using animal models. Recently, zebrafish (Danio rerio) embryos have become increasingly attractive in early drug discovery. Herein, we established a zebrafish embryo model of S. aureus infection for evaluation of in vivo efficacy of novel potential antimicrobials. A local infection was induced by microinjecting mCherry-expressing S. aureus Newman followed by treatment with reference antibiotics via microinjection into different injection sites as well as via waterborne exposure to study the impact of the administration route on efficacy. We successfully used the developed model to evaluate the in vivo activity of the natural product sorangicin A, for which common mouse models were not successful due to fast degradation in plasma. In conclusion, we present a novel screening platform for assessing in vivo activity at the antibiotic discovery stage. Furthermore, this work provides consideration for the choice of an appropriate administration route based on the physicochemical properties of tested drugs.

Physics-informed deep learning for T2-deblurred superresolution turbo spin echo MRI.

Deep learning superresolution (SR) is a promising approach to reduce MRI scan time without requiring custom sequences or iterative reconstruction. Previous deep learning SR approaches have generated low-resolution training images by simple k-space truncation, but this does not properly model in-plane turbo spin echo (TSE) MRI resolution degradation, which has variable T2 relaxation effects in different k-space regions. To fill this gap, we developed a T2 -deblurred deep learning SR method for the SR of 3D-TSE images. A SR generative adversarial network was trained using physically realistic resolution degradation (asymmetric T2 weighting of raw high-resolution k-space data). For comparison, we trained the same network structure on previous degradation models without TSE physics modeling. We tested all models for both retrospective and prospective SR with 3 × 3 acceleration factor (in the two phase-encoding directions) of genetically engineered mouse embryo model TSE-MR images. The proposed method can produce high-quality 3 × 3 SR images for a typical 500-slice volume with 6-7 mouse embryos. Because 3 × 3 SR was performed, the image acquisition time can be reduced from 15 h to 1.7 h. Compared to previous SR methods without TSE modeling, the proposed method achieved the best quantitative imaging metrics for both retrospective and prospective evaluations and achieved the best imaging-quality expert scores for prospective evaluation. The proposed T2 -deblurring method improved accuracy and image quality of deep learning-based SR of TSE MRI. This method has the potential to accelerate TSE image acquisition by a factor of up to 9.

The influence of different sugammadex doses on neural tube development in early-stage chick embryos.

Sugammadex is a modified gamma-cyclodextrin that has been developed with the goal of reversing the steroidal neuromuscular blocking agents. The aim of the present study is to investigate the effects of different sugammadex doses on embryologic and neural tube development in an early-stage chick embryo model. A total of 100 specific pathogen-free, fertilized domestic chicken eggs were randomly divided into five groups (n = 20, each), and placed in an automatic cycle incubator. The eggs in the "control (C)" group were incubated without administration of any drug till the end of the experiment. Sub-blastodermic administration of 0.9% NaCl as vehicle control (VC) and different doses of sugammadex solutions prepared with the latter [2 mg/mL (LD), 4 mg/mL (MD), 16 mg/mL (HD)] were performed at 30 hr of incubation. All embryos were removed from the eggs at 72 hr when they were expected to reach Hamburger-Hamilton (HH) stages 19-20, then they were fixed, and evaluated histo-morphologically. Embryonic development was not observed in 11 eggs (1 in C, 1 in VC; 3 in LD, 3 in MD, and 3 in HD). All the developed embryos were compatible with the HH stages 19-20. A neural tube closure defect was detected in one embryo in the HD group. No statistically significant difference was found between the groups in terms of embryonic and neural tube developments. No significant association was found between the drug and adverse outcomes; however, a trend with dosing was seen. Further studies are required before conclude on safety and extrapolate these results to human beings.

Human stem cell-derived embryo models: Toward ethically appropriate regulations and policies.

Human pluripotent stem cells can be used to derive invitro models recapitulating post-implantation human embryos. While useful for research purposes, such integrated embryo models raise ethical issues that need to be addressed to facilitate ethically appropriate policies and regulations that permit scientific ingenuity and medical progress.

Stem cell-derived embryo models: a frontier of human embryology.

Studying human development remains difficult due to limited accessibility to human embryonic tissues. Prompted by the availability of human stem cells that share molecular and cellular similarities with embryonic and extraembryonic cells in peri-implantation human embryos, researchers have now successfully developed stem cell-based human embryo models that are promising as experimental tools for studying early human development. In this Perspective, we discuss the current progress in mouse and human stem cell-derived embryo models and highlight their promising applications in advancing the fundamental understanding of mammalian development.

Seven days in the life cycle of Homo sapiens

The second week of embryonic development is a critical phase of the human life cycle and one that has been largely inaccessible to scientific investigation. Recent studies of human embryo models built from stem cells promise to yield dramatic insights into the key events of cell specification and morphogenesis that occur during this brief window of embryogenesis.