WormGUIDES is an open-source dynamic embryonic system designed to facilitate global understanding of cellular decisions in the developing nervous system of the nematode C. elegans. WormGUIDES was designed to allow inv...WormGUIDES is an open-source dynamic embryonic system designed to facilitate global understanding of cellular decisions in the developing nervous system of the nematode C. elegans. WormGUIDES was designed to allow investigation and exploration of the observational results of the C. elegans life cycle from laboratory experiments. In the process of a mechanistic C. elegans model development, some functionalities of WormGUIDES needed to be enhanced to support model validation and verification. In this study, a new way to visualize 3-dimentional vectors within WormGUIDES was investigated and presented. Then, the practical values of this method were demonstrated by visualizing two biologically significant directions (i.e., division orientation and cell polarity) of individual embryonic cells in C. elegans. Lastly, a mathematic approach was designed to illustrate the differences between these two sets of vectors and provide easy indications of the location of these individual cells that have large data discrepancies within the C. elegans embryonic system.展开更多
AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow ...AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow fluorescent protein were cultured in three-dimensional(3-D) self-assembling scaffolds and compared with traditional two-dimentional(2-D) culture techniques requiring mouse embryonic fibroblast feeder layers or leukemia inhibitory factor. 3-D scaffolds encapsulating ESCs were prepared by mixing ESCs with polyethylene glycol tetra-acrylate(PEG-4-Acr) and thiolfunctionalized dextran(Dex-SH). Distribution of ESCs in 3-D was monitored by confocal microscopy. Viability and proliferation of encapsulated cells during long-term culture were determined by propidium iodide as well as direct cell counts and PrestoB lue(PB) assays. Genetic expression of pluripotency markers(Oct4, Nanog, Klf4, and Sox2) in ESCs grown under 2-D and 3-D cultureconditions was examined by quantitative real-time polymerase chain reaction. Protein expression of selected stemness markers was determined by two different methods, immunofluorescence staining(Oct4 and Nanog) and western blot analysis(Oct4, Nanog, and Klf4). Pluripotency of 3-D scaffold grown ESCs was analyzed by in vivo teratoma assay and in vitro differentiation via embryoid bodies into cells of all three germ layers. RESULTS: Self-assembling scaffolds encapsulating ESCs for 3-D culture without the loss of cell viability were prepared by mixing PEG-4-Acr and Dex-SH(1:1 v/v) to a final concentration of 5%(w/v). Scaffold integrity was dependent on the degree of thiol substitution of Dex-SH and cell concentration. Scaffolds prepared using Dex-SH with 7.5% and 33% thiol substitution and incubated in culture medium maintained their integrity for 11 and 13 d without cells and 22 ± 5 d and 37 ± 5 d with cells, respectively. ESCs formed compact colonies, which progressively increased in size over time due to cell proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels(P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury(mesoderm), NCAM(ectoderm), and GATA4(endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation.展开更多
Innexin proteins are a class of transmembrane proteins existing in invertebrates and they have diverse biological functions. The innexin protein Sp-inx2 has been demonstrated to play roles in immune response and promo...Innexin proteins are a class of transmembrane proteins existing in invertebrates and they have diverse biological functions. The innexin protein Sp-inx2 has been demonstrated to play roles in immune response and promotion of cell apoptosis in the mud crab Scylla paramamosain . One novel innexin gene, named as Sp-inx3 was characterized from S. paramamosin in this study, with an open reading frame of 1 101 bp encoding 367 amino acid residues. Multiple sequence alignment revealed that the Sp-inx3 is highly homologous with innexin3 of Cancer boredis and Homorus americanus . Quantitative real-time PCR (qPCR) and the western blotting results revealed that Sp-inx3 gene was expressed predominantly in the eyestalk, brain, and thoracic ganglion mass in both female and male crabs. The immunohistochemistry assay (IHC) also showed the widespread and intense immunoreactivity of Sp-inx3 in the brain and thoracic ganglion mass. Sp-inx3 mRNA transcription profi les exhibited signifi cantly higher expression from the embryo1 to embryo4 period and low level of expression at the prehatching period and zoea I larva period of S . paramamosain . These results indicate that the Sp-inx3 may play an important role in the nervous system and early embryonic development of S . paramamosain.展开更多
With the development of high-throughput sequencing technology in the post-genomic era, researchers have concentrated their efforts on elucidating the relationships between genes and their corresponding functions. Rece...With the development of high-throughput sequencing technology in the post-genomic era, researchers have concentrated their efforts on elucidating the relationships between genes and their corresponding functions. Recently, important progress has been achieved in the generation of genetically modified mice based on CRISPR/Cas9 and haploid embryonic stem cell （haESC） approaches, which provide new platforms for gene function analysis, human disease modeling, and gene therapy. Here, we review the CRISPR/Cas9 and haESC technology for the generation of genetically modified mice and discuss the key challenges in the application of these approaches.展开更多
In an experiment in rats with electrolytic lesion of the compact part of substantia nigra (SN) and after allotransplantation of the embryonic tissue of SN in the caudate nucleus the features of movement and emotional ...In an experiment in rats with electrolytic lesion of the compact part of substantia nigra (SN) and after allotransplantation of the embryonic tissue of SN in the caudate nucleus the features of movement and emotional behavior in the Open Field Test (OFT), the rotation movements caused by an administration of amphetamine, a content of catecholamines in the caudate nucleus, hypothalamus and blood plasma have been investigated. It is shown that the electrolytic lesion causes violations of the statokinetic reflexes, the horizontal and the vertical movement activity, enhances the rotatory behavior, slow the orienttate-searching and the emotional reactions that combined with disbalance in dopamine-and noradrenalinetransmitter systems functioning. Allotransplantation of the embryonic dofaminsynthesizing brain tissue contributes to the restoration of movement activeity and its specific neurotransmitter ensuring.展开更多
Human cytomegalovirus(HCMV)poses a significant risk of neural damage during pregnancy.As the most prevalent intrauterine infectious agent in low-and middle-income countries,HCMV disrupts the development of neural stem...Human cytomegalovirus(HCMV)poses a significant risk of neural damage during pregnancy.As the most prevalent intrauterine infectious agent in low-and middle-income countries,HCMV disrupts the development of neural stem cells,leading to fetal malformations and abnormal structural and physiological functions in the fetal brain.This review summarizes the current understanding of how HCMV infection dysregulates the Wnt signaling pathway to induce fetal malformations and discusses current management strategies.展开更多
Stem cell-based brain repair is a promising emergent therapy for Parkinson's disease based on years of foundational research using human fetal donors as a cell source.Unlike current therapeutic options for patient...Stem cell-based brain repair is a promising emergent therapy for Parkinson's disease based on years of foundational research using human fetal donors as a cell source.Unlike current therapeutic options for patients,this approach has the potential to provide longterm stem cell–derived reconstruction and restoration of the dopaminergic input to denervated regions of the brain allowing for restoration of certain functions to patients.The ultimate clinical success of stem cell–derived brain repair will depend on both the safety and efficacy of the approach and the latter is dependent on the ability of the transplanted cells to survive and differentiate into functional dopaminergic neurons in the Parkinsonian brain.Because the pre-clinical literature suggests that there is considerable variability in survival and differentiation between studies,the aim of this systematic review was to assess these parameters in human stem cell-derived dopaminergic progenitor transplant studies in animal models of Parkinson's disease.A defined systematic search of the PubMed database was completed to identify relevant studies published up to March 2024.After screening,76 articles were included in the analysis from which 178 separate transplant studies were identified.From these,graft survival could be assessed in 52 studies and differentiation in 129 studies.Overall,we found that graft survival ranged from<1% to 500% of cells transplanted,with a median of 51%of transplanted cells surviving in the brain;while dopaminergic differentiation of the cells ranged from 0% to 46% of cells transplanted with a median of 3%.This systematic review suggests that there is considerable scope for improvement in the differentiation of stem cell-derived dopaminergic progenitors to maximize the therapeutic potential of this approach for patients.展开更多
Spermatozoa have a highly complex RNA profile.Several of these transcripts are suggested as biomarkers for male infertility and contribute to early development.To analyze the differences between sperm RNA quantity and...Spermatozoa have a highly complex RNA profile.Several of these transcripts are suggested as biomarkers for male infertility and contribute to early development.To analyze the differences between sperm RNA quantity and expression of protamine(PRM1 and PRM2)and testis-specific histone 2B(TH2B)genes,spermatozoa from 33 patients who enrolled in assisted reproduction treatment(ART)program were analyzed.Sperm RNA of teratozoospermic(T),oligoteratozoospermic(OT),and normozoospermic(N)samples was extracted,and the differences in transcript levels among the study groups were analyzed by quantitative real-time polymerase chain reaction(qRT-PCR).The correlations of total RNA per spermatozoon and the expression of the transcripts were evaluated in relation to sperm characteristics and preimplantation embryo development.The mean(±standard deviation)RNA amount per spermatozoon was 28.48(±23.03)femtogram in the overall group and was significantly higher in the OT group than that in N and T groups.Total sperm RNA and gene expression of PRM1 and PRM2 genes were related to preimplantation embryo development and developmental arrest.Specific sperm characteristics were correlated with the expressions of PRM1,PRM2,or TH2B genes.We conclude that the sperm RNA amount and composition are important factors and might influence early embryonic development and also differ in different cases of male infertility.展开更多
The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can ...The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can cause reduced fertil-ity and reproductive disorders in mammals.Nevertheless,the local impact of elevated stress hormones on the ovi-duct epithelium has received limited attention to date,except for a few reports on polyovulatory species like mice and pigs.In this study,we focused on the effects of chronic maternal stress on cattle,given its association with infertil-ity issues in this monoovulatory species.Bovine oviduct epithelial cells(BOEC)differentiated at the air–liquid interface(ALI)were stimulated with 250 nmol/L cortisol for 1 or 3 weeks.Subsequently,they were assessed for morphology,bioelectrical properties,and gene expression related to oviduct function,glucocorticoid pathway,cortisol metabo-lism,inflammation,and apoptosis.Results revealed adverse effects of cortisol on epithelium structure,featured by deciliation,vacuole formation,and multilayering.Additionally,cortisol exposure led to an increase in transepithelial potential difference,downregulated mRNA expression of the major glucocorticoid receptor(NR3C1),upregulated the expression of cortisol-responsive genes(FKBP5,TSC22D3),and significant downregulation of oviductal glycopro-tein 1(OVGP1)and steroid receptors PGR and ESR1.The systematic comparison to a similar experiment previously performed by us in porcine oviduct epithelial cells,indicated that bovine cultures were more susceptible to elevated cortisol levels than porcine.The distinct responses between both species are likely linked to their divergence in the cortisol-induced expression changes of HSD11B2,an enzyme controlling the cellular capacity to metabolise cortisol.These findings provide insights into the species-specific reactions and reproductive consequences triggered by maternal stress.展开更多
Microglia have been recognized as a unique cell population in the central nervous system(CNS)for more than a century[1].However,it was not until 2010 that their developmental origin was clarified.Rather than arising f...Microglia have been recognized as a unique cell population in the central nervous system(CNS)for more than a century[1].However,it was not until 2010 that their developmental origin was clarified.Rather than arising from the neuroectoderm,microglia are derived from erythromyeloid progenitors in the embryonic yolk sac[2].展开更多
Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our publ...Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our published work,we request the publication of a corrigendum with the corrected image.We apologize for this oversight and any confusion it may have caused.The amended figure is provided in the updated Supplementary Materials.展开更多
Background Early embryo development plays a pivotal role in determining pregnancy outcomes,postnatal development,and lifelong health.Therefore,the strategic selection of functional nutrients to enhance embryo developm...Background Early embryo development plays a pivotal role in determining pregnancy outcomes,postnatal development,and lifelong health.Therefore,the strategic selection of functional nutrients to enhance embryo development is of paramount importance.In this study,we established a stable porcine trophectoderm cell line expressing dual fluorescent reporter genes driven by the CDX2 and TEAD4 gene promoter segments using lentiviral transfection.Results Three amino acid metabolites—kynurenic acid,taurine,and tryptamine—met the minimum z-score criteria of 2.0 for both luciferase and Renilla luciferase activities and were initially identified as potential metabolites for embryo development,with their beneficial effects validated by qPCR.Given that the identified metabolites are closely related to methionine,arginine,and tryptophan,we selected these three amino acids,using lysine as a standard,and employed response surface methodology combined with our high-throughput screening cell model to efficiently screen and optimize amino acid combination conducive to early embryo development.The optimized candidate amino acid system included lysine(1.87 mmol/L),methionine(0.82 mmol/L),tryptophan(0.23 mmol/L),and arginine(3 mmol/L),with the ratio of 1:0.43:0.12:1.60.In vitro experiments confirmed that this amino acid system enhances the expression of key genes involved in early embryonic development and improves in vitro embryo adhesion.Transcriptomic analysis of blastocysts suggested that candidate amino acid system enhances early embryo development by regulating early embryonic cell cycle and differentiation,as well as improving nutrient absorption.Furthermore,based on response surface methodology,400 sows were used to verify this amino acid system,substituting arginine with the more cost-effective N-carbamoyl glutamate(NCG),a precursor of arginine.The optimal dietary amino acid requirement was predicted to be 0.71%lysine,0.32%methionine,0.22%tryptophan,and 0.10%NCG for sows during early gestation.The optimized amino acid system ratio of the feed,derived from the peripheral release of essential amino acids,was found to be 1:0.45:0.13,which is largely consistent with the results obtained from the cell model optimization.Subsequently,we furtherly verified that this optimal dietary amino acid system significantly increased total litter size,live litter size and litter weight in sows.Conclusions In summary,we successfully established a dual-fluorescent high-throughput screening cell model for the efficient identification of potential nutrients that would promote embryo development and implantation.This innovative approach overcomes the limitations of traditional amino acid nutrition studies in sows,providing a more effective model for enhancing reproductive outcomes.展开更多
Spatial transcriptomics technology provides novel insights into the spatial organization of gene expression during embryonic development.In this study,we propose a method that integrates analysis across both temporal ...Spatial transcriptomics technology provides novel insights into the spatial organization of gene expression during embryonic development.In this study,we propose a method that integrates analysis across both temporal and spatial dimensions to investigate spatial transcriptomics data from mouse embryos at different developmental stages.We quantified the spatial expression pattern of each gene at various stages by calculating its Moran’s I.Furthermore,by employing time-series clustering to identify dynamic co-expression modules,we identified several developmentally stage-specific regulatory gene modules.A key finding was the presence of distinct,stage-specific gene network modules across different developmental periods:Early modules focused on morphogenesis,mid-stage on organ development,and late-stage on neural and tissue maturation.Functional enrichment analysis further confirmed the core biological functions of each module.The dynamic,spatially-resolved gene expression model constructed in this study not only provides new biological insights into the programmed spatiotemporal reorganization of gene regulatory networks during embryonic development but also presents an effective approach for analyzing complex spatiotemporal omics data.This work provides a new perspective for understanding developmental biology,regenerative medicine,and related fields.展开更多
Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response.Stem cell therapy can be a promising treatment strategy for periodontitis,but the relevant mechanism is complica...Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response.Stem cell therapy can be a promising treatment strategy for periodontitis,but the relevant mechanism is complicated.This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells(hESC-MSCs)for the treatment of periodontitis.The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure.Human gingival fibroblasts(HGFs)were exposed to 5μg/mL lipopolysaccharide(LPS)for 24 h to establish a cell injury model.When treated with hESC-MSCs or mitochondria derived from hESC-MSCs,HGFs showed reduced expression of inflammatory genes,increased adenosine triphosphate(ATP)level,decreased reactive oxygen species(ROS)production,and enhanced mitochondrial function compared to the control.The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%.Besides,a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression,as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues.In conclusion,our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs,suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.展开更多
We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota （Amphibia, Anura, Ranidae）. Embryos were derived from artificial fertilization of frogs’ eggs, and the...We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota （Amphibia, Anura, Ranidae）. Embryos were derived from artificial fertilization of frogs’ eggs, and the staging of development was based on morphological and physiological characteristics. Two major periods of development were designated： i） early embryonic period, from fertilization to operculum completion stage, lasted for 324 h at water temperature （WT） 18 ？23℃; ii） larval period, from operculum completion stage to tail absorbed stage, took 1207 h at WT 20 ？ 24℃. Tadpoles of the concave-eared torrent frogs showed no evidence of abdominal sucker. Absence of this key characteristic supports the view from molecular systematics that concave-eared torrent frog does not belong to the genus Amolops. Two cleavage patterns were observed in embryos at 8-cell and 16-cell stages, with Pattern I2 （latitudinal cleavage at the 8-cell stage, and meridional cleavage at the 16-cell stage with two perpendicular meridional furrows） being the predominant pattern and only 1.5% belonging to Pattern II （meridional cleavage at the 8-cell stage and latitudinal cleavage at the 16-cell stage）. The factors affecting cleavage and hatching ratios, developmental speed, and ecological adaptation were discussed.展开更多
Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,wh...Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm（DE） cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins（CK7,CK18,CK19,CK20,and OV-6） and genes（GSTPi,IB4,and HNF1β）.At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ～90% to ～20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.展开更多
As aerospace vehicles travel in a hellish environment,the reliability of the measuring and controlling systems has played a critical role in the credibility of a whole airborne system.Embryo-electronic system is a bio...As aerospace vehicles travel in a hellish environment,the reliability of the measuring and controlling systems has played a critical role in the credibility of a whole airborne system.Embryo-electronic system is a bionic hardware capable of self-diagnosing and self-healing.This article presents a new approach to design embryo-electronic systems and introduces their bionic principles,system structures and fanlt-tolerant mechanism.As the current methods cannot meet the requirements for large-scale embryo-electronic systems,this article advances a new shift-register-based configuration memory of embryonic system to solve the problem by using the inter-cell communication to reduce the gene storage capacity of a single cell.The article designs an overall structure of the shift-register-based configuration memories of the embryonic system and connects them into a chain structure.The article also designs an inner circuit of the cell,the control of shift-register-based configuration memory and the way of runtime dynamic configuration.The simulation of field programmable gate array(FPGA)evidences the realizability of the proposed design.Compared to the SRAM-based one,this memory can save 90%of the area when constructing embryonic systems larger than 128×128 under the same condition.展开更多
[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for i...[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for intensive breeding and propagation by artificial insemination. The entire embryonic development process of L. crassilabris from insemination to hatching out of larvae fish was observed consecutively. [Result] Fertilized eggs of L. crassilabris are spherical, and the egg diameter is about 2.2-2.4 mm after water absorption; under conditions of water temperature ranging from 27 to 28 ℃, the embryonic development of L. crassilabris from insemination to hatching out of larvae fish lasted 2 496 min, with a total accumulated temperature of 1 123-1 165 h·℃. [Conclusion] This study is advantageous to better understand the characteristics of embryonic development of L. crassilabris and provides basic biological data for protection and utilization of fish resources and other related work.展开更多
[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with te...[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5. The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage. The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] It provided important reference for studying artificial propagation and genetic breeding of loach.展开更多
[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down ...[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.展开更多
文摘WormGUIDES is an open-source dynamic embryonic system designed to facilitate global understanding of cellular decisions in the developing nervous system of the nematode C. elegans. WormGUIDES was designed to allow investigation and exploration of the observational results of the C. elegans life cycle from laboratory experiments. In the process of a mechanistic C. elegans model development, some functionalities of WormGUIDES needed to be enhanced to support model validation and verification. In this study, a new way to visualize 3-dimentional vectors within WormGUIDES was investigated and presented. Then, the practical values of this method were demonstrated by visualizing two biologically significant directions (i.e., division orientation and cell polarity) of individual embryonic cells in C. elegans. Lastly, a mathematic approach was designed to illustrate the differences between these two sets of vectors and provide easy indications of the location of these individual cells that have large data discrepancies within the C. elegans embryonic system.
基金Oakland University and Oakland University-William Beaumont Institute for Stem Cell and Regenerative Medicine(OU-WB ISCRM)
文摘AIM: To devise a simplified and efficient method for long-term culture and maintenance of embryonic stem cells requiring less frequent passaging. METHODS: Mouse embryonic stem cells(ESCs) labeled with enhanced yellow fluorescent protein were cultured in three-dimensional(3-D) self-assembling scaffolds and compared with traditional two-dimentional(2-D) culture techniques requiring mouse embryonic fibroblast feeder layers or leukemia inhibitory factor. 3-D scaffolds encapsulating ESCs were prepared by mixing ESCs with polyethylene glycol tetra-acrylate(PEG-4-Acr) and thiolfunctionalized dextran(Dex-SH). Distribution of ESCs in 3-D was monitored by confocal microscopy. Viability and proliferation of encapsulated cells during long-term culture were determined by propidium iodide as well as direct cell counts and PrestoB lue(PB) assays. Genetic expression of pluripotency markers(Oct4, Nanog, Klf4, and Sox2) in ESCs grown under 2-D and 3-D cultureconditions was examined by quantitative real-time polymerase chain reaction. Protein expression of selected stemness markers was determined by two different methods, immunofluorescence staining(Oct4 and Nanog) and western blot analysis(Oct4, Nanog, and Klf4). Pluripotency of 3-D scaffold grown ESCs was analyzed by in vivo teratoma assay and in vitro differentiation via embryoid bodies into cells of all three germ layers. RESULTS: Self-assembling scaffolds encapsulating ESCs for 3-D culture without the loss of cell viability were prepared by mixing PEG-4-Acr and Dex-SH(1:1 v/v) to a final concentration of 5%(w/v). Scaffold integrity was dependent on the degree of thiol substitution of Dex-SH and cell concentration. Scaffolds prepared using Dex-SH with 7.5% and 33% thiol substitution and incubated in culture medium maintained their integrity for 11 and 13 d without cells and 22 ± 5 d and 37 ± 5 d with cells, respectively. ESCs formed compact colonies, which progressively increased in size over time due to cell proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels(P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury(mesoderm), NCAM(ectoderm), and GATA4(endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation.
基金Supported by the Fundamental Research Funds for the Central Universities(No.20720180100)the National Natural Science Foundation of China(NSFC)(Nos.U1205123,41676158)the Fujian Science and Technology Department(No.2014N2004)
文摘Innexin proteins are a class of transmembrane proteins existing in invertebrates and they have diverse biological functions. The innexin protein Sp-inx2 has been demonstrated to play roles in immune response and promotion of cell apoptosis in the mud crab Scylla paramamosain . One novel innexin gene, named as Sp-inx3 was characterized from S. paramamosin in this study, with an open reading frame of 1 101 bp encoding 367 amino acid residues. Multiple sequence alignment revealed that the Sp-inx3 is highly homologous with innexin3 of Cancer boredis and Homorus americanus . Quantitative real-time PCR (qPCR) and the western blotting results revealed that Sp-inx3 gene was expressed predominantly in the eyestalk, brain, and thoracic ganglion mass in both female and male crabs. The immunohistochemistry assay (IHC) also showed the widespread and intense immunoreactivity of Sp-inx3 in the brain and thoracic ganglion mass. Sp-inx3 mRNA transcription profi les exhibited signifi cantly higher expression from the embryo1 to embryo4 period and low level of expression at the prehatching period and zoea I larva period of S . paramamosain . These results indicate that the Sp-inx3 may play an important role in the nervous system and early embryonic development of S . paramamosain.
基金supported by the National Natural Science Foundation of China(3731530048C1202)
文摘With the development of high-throughput sequencing technology in the post-genomic era, researchers have concentrated their efforts on elucidating the relationships between genes and their corresponding functions. Recently, important progress has been achieved in the generation of genetically modified mice based on CRISPR/Cas9 and haploid embryonic stem cell （haESC） approaches, which provide new platforms for gene function analysis, human disease modeling, and gene therapy. Here, we review the CRISPR/Cas9 and haESC technology for the generation of genetically modified mice and discuss the key challenges in the application of these approaches.
文摘In an experiment in rats with electrolytic lesion of the compact part of substantia nigra (SN) and after allotransplantation of the embryonic tissue of SN in the caudate nucleus the features of movement and emotional behavior in the Open Field Test (OFT), the rotation movements caused by an administration of amphetamine, a content of catecholamines in the caudate nucleus, hypothalamus and blood plasma have been investigated. It is shown that the electrolytic lesion causes violations of the statokinetic reflexes, the horizontal and the vertical movement activity, enhances the rotatory behavior, slow the orienttate-searching and the emotional reactions that combined with disbalance in dopamine-and noradrenalinetransmitter systems functioning. Allotransplantation of the embryonic dofaminsynthesizing brain tissue contributes to the restoration of movement activeity and its specific neurotransmitter ensuring.
基金supported by the Natural Science Foundation of Shandong Province(ZR2019MC059)the Traditional Chinese Medicine Science Project of Shandong Province(M-2023093)the Weifang Municipal Science and Technology Development Program(2025YX037).
文摘Human cytomegalovirus(HCMV)poses a significant risk of neural damage during pregnancy.As the most prevalent intrauterine infectious agent in low-and middle-income countries,HCMV disrupts the development of neural stem cells,leading to fetal malformations and abnormal structural and physiological functions in the fetal brain.This review summarizes the current understanding of how HCMV infection dysregulates the Wnt signaling pathway to induce fetal malformations and discusses current management strategies.
基金supported by research grants from the Michael J Fox Foundation for Parkinson’s Research(grant numbers:17244 and 023410)Science Foundation Ireland(Grant Numbers:19/FFP/6554)(to ED)。
文摘Stem cell-based brain repair is a promising emergent therapy for Parkinson's disease based on years of foundational research using human fetal donors as a cell source.Unlike current therapeutic options for patients,this approach has the potential to provide longterm stem cell–derived reconstruction and restoration of the dopaminergic input to denervated regions of the brain allowing for restoration of certain functions to patients.The ultimate clinical success of stem cell–derived brain repair will depend on both the safety and efficacy of the approach and the latter is dependent on the ability of the transplanted cells to survive and differentiate into functional dopaminergic neurons in the Parkinsonian brain.Because the pre-clinical literature suggests that there is considerable variability in survival and differentiation between studies,the aim of this systematic review was to assess these parameters in human stem cell-derived dopaminergic progenitor transplant studies in animal models of Parkinson's disease.A defined systematic search of the PubMed database was completed to identify relevant studies published up to March 2024.After screening,76 articles were included in the analysis from which 178 separate transplant studies were identified.From these,graft survival could be assessed in 52 studies and differentiation in 129 studies.Overall,we found that graft survival ranged from<1% to 500% of cells transplanted,with a median of 51%of transplanted cells surviving in the brain;while dopaminergic differentiation of the cells ranged from 0% to 46% of cells transplanted with a median of 3%.This systematic review suggests that there is considerable scope for improvement in the differentiation of stem cell-derived dopaminergic progenitors to maximize the therapeutic potential of this approach for patients.
基金supported by the Scientific Research Projects Coordination Unit of Istanbul University(No.13930).
文摘Spermatozoa have a highly complex RNA profile.Several of these transcripts are suggested as biomarkers for male infertility and contribute to early development.To analyze the differences between sperm RNA quantity and expression of protamine(PRM1 and PRM2)and testis-specific histone 2B(TH2B)genes,spermatozoa from 33 patients who enrolled in assisted reproduction treatment(ART)program were analyzed.Sperm RNA of teratozoospermic(T),oligoteratozoospermic(OT),and normozoospermic(N)samples was extracted,and the differences in transcript levels among the study groups were analyzed by quantitative real-time polymerase chain reaction(qRT-PCR).The correlations of total RNA per spermatozoon and the expression of the transcripts were evaluated in relation to sperm characteristics and preimplantation embryo development.The mean(±standard deviation)RNA amount per spermatozoon was 28.48(±23.03)femtogram in the overall group and was significantly higher in the OT group than that in N and T groups.Total sperm RNA and gene expression of PRM1 and PRM2 genes were related to preimplantation embryo development and developmental arrest.Specific sperm characteristics were correlated with the expressions of PRM1,PRM2,or TH2B genes.We conclude that the sperm RNA amount and composition are important factors and might influence early embryonic development and also differ in different cases of male infertility.
基金German research Foundation(DFG,grant numbers:CH2321/1–1 and SCHO1231/7–1)JH has received a scholarship from the Chinese Scholarship Council(CSC No.:201908350115).
文摘The oviduct epithelium is the initial maternal contact site for embryos after fertilization,offering the microenviron-ment before implantation.This early gestation period is particularly sensitive to stress,which can cause reduced fertil-ity and reproductive disorders in mammals.Nevertheless,the local impact of elevated stress hormones on the ovi-duct epithelium has received limited attention to date,except for a few reports on polyovulatory species like mice and pigs.In this study,we focused on the effects of chronic maternal stress on cattle,given its association with infertil-ity issues in this monoovulatory species.Bovine oviduct epithelial cells(BOEC)differentiated at the air–liquid interface(ALI)were stimulated with 250 nmol/L cortisol for 1 or 3 weeks.Subsequently,they were assessed for morphology,bioelectrical properties,and gene expression related to oviduct function,glucocorticoid pathway,cortisol metabo-lism,inflammation,and apoptosis.Results revealed adverse effects of cortisol on epithelium structure,featured by deciliation,vacuole formation,and multilayering.Additionally,cortisol exposure led to an increase in transepithelial potential difference,downregulated mRNA expression of the major glucocorticoid receptor(NR3C1),upregulated the expression of cortisol-responsive genes(FKBP5,TSC22D3),and significant downregulation of oviductal glycopro-tein 1(OVGP1)and steroid receptors PGR and ESR1.The systematic comparison to a similar experiment previously performed by us in porcine oviduct epithelial cells,indicated that bovine cultures were more susceptible to elevated cortisol levels than porcine.The distinct responses between both species are likely linked to their divergence in the cortisol-induced expression changes of HSD11B2,an enzyme controlling the cellular capacity to metabolise cortisol.These findings provide insights into the species-specific reactions and reproductive consequences triggered by maternal stress.
基金supported in part by National Science and Technology Major Project of National Health Commission of China(2023ZD0520300)the National Natural Science Foundation of China(32130037 and 32370953)the Tsinghua University Initiative Scientific Research Program,SXMU-Tsinghua Collaborative Innovation Center for Frontier Medicine and the Research Fund of Vanke School of Public Health,Tsinghua University.H.Q.is a New Cornerstone Investigator.
文摘Microglia have been recognized as a unique cell population in the central nervous system(CNS)for more than a century[1].However,it was not until 2010 that their developmental origin was clarified.Rather than arising from the neuroectoderm,microglia are derived from erythromyeloid progenitors in the embryonic yolk sac[2].
文摘Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our published work,we request the publication of a corrigendum with the corrected image.We apologize for this oversight and any confusion it may have caused.The amended figure is provided in the updated Supplementary Materials.
基金supported by National Natural Science Foundation of China (32172747 and 32425052)
文摘Background Early embryo development plays a pivotal role in determining pregnancy outcomes,postnatal development,and lifelong health.Therefore,the strategic selection of functional nutrients to enhance embryo development is of paramount importance.In this study,we established a stable porcine trophectoderm cell line expressing dual fluorescent reporter genes driven by the CDX2 and TEAD4 gene promoter segments using lentiviral transfection.Results Three amino acid metabolites—kynurenic acid,taurine,and tryptamine—met the minimum z-score criteria of 2.0 for both luciferase and Renilla luciferase activities and were initially identified as potential metabolites for embryo development,with their beneficial effects validated by qPCR.Given that the identified metabolites are closely related to methionine,arginine,and tryptophan,we selected these three amino acids,using lysine as a standard,and employed response surface methodology combined with our high-throughput screening cell model to efficiently screen and optimize amino acid combination conducive to early embryo development.The optimized candidate amino acid system included lysine(1.87 mmol/L),methionine(0.82 mmol/L),tryptophan(0.23 mmol/L),and arginine(3 mmol/L),with the ratio of 1:0.43:0.12:1.60.In vitro experiments confirmed that this amino acid system enhances the expression of key genes involved in early embryonic development and improves in vitro embryo adhesion.Transcriptomic analysis of blastocysts suggested that candidate amino acid system enhances early embryo development by regulating early embryonic cell cycle and differentiation,as well as improving nutrient absorption.Furthermore,based on response surface methodology,400 sows were used to verify this amino acid system,substituting arginine with the more cost-effective N-carbamoyl glutamate(NCG),a precursor of arginine.The optimal dietary amino acid requirement was predicted to be 0.71%lysine,0.32%methionine,0.22%tryptophan,and 0.10%NCG for sows during early gestation.The optimized amino acid system ratio of the feed,derived from the peripheral release of essential amino acids,was found to be 1:0.45:0.13,which is largely consistent with the results obtained from the cell model optimization.Subsequently,we furtherly verified that this optimal dietary amino acid system significantly increased total litter size,live litter size and litter weight in sows.Conclusions In summary,we successfully established a dual-fluorescent high-throughput screening cell model for the efficient identification of potential nutrients that would promote embryo development and implantation.This innovative approach overcomes the limitations of traditional amino acid nutrition studies in sows,providing a more effective model for enhancing reproductive outcomes.
基金supported by the National Natural Science Foundation of China(Grant Nos.12090052,U24A2014,and 12325405).
文摘Spatial transcriptomics technology provides novel insights into the spatial organization of gene expression during embryonic development.In this study,we propose a method that integrates analysis across both temporal and spatial dimensions to investigate spatial transcriptomics data from mouse embryos at different developmental stages.We quantified the spatial expression pattern of each gene at various stages by calculating its Moran’s I.Furthermore,by employing time-series clustering to identify dynamic co-expression modules,we identified several developmentally stage-specific regulatory gene modules.A key finding was the presence of distinct,stage-specific gene network modules across different developmental periods:Early modules focused on morphogenesis,mid-stage on organ development,and late-stage on neural and tissue maturation.Functional enrichment analysis further confirmed the core biological functions of each module.The dynamic,spatially-resolved gene expression model constructed in this study not only provides new biological insights into the programmed spatiotemporal reorganization of gene regulatory networks during embryonic development but also presents an effective approach for analyzing complex spatiotemporal omics data.This work provides a new perspective for understanding developmental biology,regenerative medicine,and related fields.
基金supported by the Key R&D Program of Zhejiang(No.2023C03072)the Medical and Health Science and Technology Program of Zhejiang Province(No.2021PY007)and the National Key Research and Development Program of China(Nos.2021YFA1301100 and 2021YFA1301101).
文摘Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response.Stem cell therapy can be a promising treatment strategy for periodontitis,but the relevant mechanism is complicated.This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells(hESC-MSCs)for the treatment of periodontitis.The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure.Human gingival fibroblasts(HGFs)were exposed to 5μg/mL lipopolysaccharide(LPS)for 24 h to establish a cell injury model.When treated with hESC-MSCs or mitochondria derived from hESC-MSCs,HGFs showed reduced expression of inflammatory genes,increased adenosine triphosphate(ATP)level,decreased reactive oxygen species(ROS)production,and enhanced mitochondrial function compared to the control.The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%.Besides,a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression,as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues.In conclusion,our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs,suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.
基金supported by the National Natural Science Fundation of China(30730029)
文摘We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota （Amphibia, Anura, Ranidae）. Embryos were derived from artificial fertilization of frogs’ eggs, and the staging of development was based on morphological and physiological characteristics. Two major periods of development were designated： i） early embryonic period, from fertilization to operculum completion stage, lasted for 324 h at water temperature （WT） 18 ？23℃; ii） larval period, from operculum completion stage to tail absorbed stage, took 1207 h at WT 20 ？ 24℃. Tadpoles of the concave-eared torrent frogs showed no evidence of abdominal sucker. Absence of this key characteristic supports the view from molecular systematics that concave-eared torrent frog does not belong to the genus Amolops. Two cleavage patterns were observed in embryos at 8-cell and 16-cell stages, with Pattern I2 （latitudinal cleavage at the 8-cell stage, and meridional cleavage at the 16-cell stage with two perpendicular meridional furrows） being the predominant pattern and only 1.5% belonging to Pattern II （meridional cleavage at the 8-cell stage and latitudinal cleavage at the 16-cell stage）. The factors affecting cleavage and hatching ratios, developmental speed, and ecological adaptation were discussed.
基金supported by research grants from Zhejiang Natural Sciences Foundation of China (Y2110911 Y2080996)the National Key Technologies R&D Program of China (2007CB947701)
文摘Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm（DE） cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins（CK7,CK18,CK19,CK20,and OV-6） and genes（GSTPi,IB4,and HNF1β）.At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ～90% to ～20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.
基金National Natural Science Foundation of China(90505013)
文摘As aerospace vehicles travel in a hellish environment,the reliability of the measuring and controlling systems has played a critical role in the credibility of a whole airborne system.Embryo-electronic system is a bionic hardware capable of self-diagnosing and self-healing.This article presents a new approach to design embryo-electronic systems and introduces their bionic principles,system structures and fanlt-tolerant mechanism.As the current methods cannot meet the requirements for large-scale embryo-electronic systems,this article advances a new shift-register-based configuration memory of embryonic system to solve the problem by using the inter-cell communication to reduce the gene storage capacity of a single cell.The article designs an overall structure of the shift-register-based configuration memories of the embryonic system and connects them into a chain structure.The article also designs an inner circuit of the cell,the control of shift-register-based configuration memory and the way of runtime dynamic configuration.The simulation of field programmable gate array(FPGA)evidences the realizability of the proposed design.Compared to the SRAM-based one,this memory can save 90%of the area when constructing embryonic systems larger than 128×128 under the same condition.
基金Supported by Science and Technology Support Program of Science and Technology Department of Jiangsu Province(BE2009335)~~
文摘[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for intensive breeding and propagation by artificial insemination. The entire embryonic development process of L. crassilabris from insemination to hatching out of larvae fish was observed consecutively. [Result] Fertilized eggs of L. crassilabris are spherical, and the egg diameter is about 2.2-2.4 mm after water absorption; under conditions of water temperature ranging from 27 to 28 ℃, the embryonic development of L. crassilabris from insemination to hatching out of larvae fish lasted 2 496 min, with a total accumulated temperature of 1 123-1 165 h·℃. [Conclusion] This study is advantageous to better understand the characteristics of embryonic development of L. crassilabris and provides basic biological data for protection and utilization of fish resources and other related work.
文摘[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5. The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage. The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] It provided important reference for studying artificial propagation and genetic breeding of loach.
基金Supported by Project of Baotou University(BSY2010-23)~~
文摘[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.
