PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “embryo model”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 91 records · Page 5Linked to original sources

Production of erythropoietic colony-forming units and erythrocytes during chick embryo development: an attempt at modelization of chick embryo erythropoiesis.

The enumeration of erythropoietic colony-forming cells in vitro has allowed us to complete previous data on changes in the various erythroid cell populations during chick embryo-genesis. Erythrocytic colony-forming units in culture (CFU-cE) which are sensitive to avian erythropoietin appear in the blastoderm as soon as the 24th hour of development. They represent most likely precursors of the megalocytic erythropoiesis, and do not seem to derive from stem cells common with normocytic erythropoiesis. Data concerning vitelline normocytic erythropoiesis were analysed in a kinetic model based on stochastic change of the stem cells. From this model it appears that 17-20 cell divisions are required for differentiation of erythrocytes from stem cells.

Animals↗

Neuronal defects in genotyped dominant megacolon (Dom) mouse embryos, a model for Hirschsprung disease.

Dominant megacolon (Dom) is one of four mutations in the mouse which can produce a phenotype similar to Hirschsprung disease in man. Here, we report that it is possible to take advantage of two microsatellite markers to genotype Dom embryos and to study enteric neuronal development in Dom embryos using whole-mount immunohistochemistry. Dom embryos present a variable defect in the ileo-caecal region, as do embryos of other murine models of Hirschsprung disease.

Animals↗

Development of a green fluorescent protein metastatic-cancer chick-embryo drug-screen model.

The chick-embryo model has been an important tool to study tumor growth, metastasis, and angiogenesis. However, an imageable model with a genetic fluorescent tag in the growing and spreading cancer cells that is stable over time has not been developed. We report here the development of such an imageable fluorescent chick-embryo metastatic cancer model with the use of green fluorescent protein (GFP). Lewis lung carcinoma cells, stably expressing GFP, were injected on the 12th day of incubation in the chick embryo. GFP-Lewis lung carcinoma metastases were visualized by fluorescence, after seven days additional incubation, in the brain, heart, and sternum of the developing chick embryo, with the most frequent site being the brain. The combination of streptokinase and gemcitabine was evaluated in this GFP metastatic model. Twelve-day-old chick embryos were injected intravenously with GFP-Lewis lung cancer cells, along with these two agents either alone or in combination. The streptokinase-gemcitabine combination inhibited metastases at all sites. The effective dose of gemcitabine was found to be 10 mg/kg and streptokinase 2000 IU per embryo. The data in this report suggest that this new stably fluorescent imageable metastatic-cancer chick-embryo model will enable rapid screening of new antimetastatic agents.

Animals↗

Chick embryo, a model to study the lethal activity of pertussis toxin, infectivity of Bordetella pertussis, and their neutralization by immune sera.

The toxin activity of Bordetella strains and acellular pertussis components was evaluated in chick embryos. Eleven-day-old embryos were found to be most suitable for determination of LD50 values. Eight of eight Bordetella pertussis strains possessed LD50 values of 10(4) to 10(6) colony-forming units per dose of 100 microL. Embryos were resistant to Bordetella parapertussis and Bordetella bronchiseptica at doses of 10(10) colony-forming units. Bacterial growth did not occur in the bloodstream or tissues of the heart, kidney, brain, liver, or lungs. Pertussis toxin activity, determined by clustering of Chinese hamster ovary cells, was found in the allantoic fluid following inoculation of virulent bacteria. Purified pertussis toxin had an LD50 of 1-2 micrograms protein/dose. Striking pathological effects of liver necrosis and oedema of the head and neck were similar following bacterial or toxin inoculation. Widespread changes occurred in the liver at the perivascular level, suggesting that after allantoic inoculation of virulent Bordetella strains, toxin is secreted, carried in the bloodstream, and causes the resultant pathology. Purified B. pertussis lipopolysaccharide was nontoxic for embryos at doses containing 30 micrograms; filamentous haemagglutinin was nontoxic at doses up to 10 micrograms. A correlation exists between the ability of serum to neutralize killing of chick embryos by toxin and to neutralize clustering of Chinese hamster ovary cells by toxin. Although the chick embryo serum neutralization studies confirm observations of mouse intracerebral infection studies that antibody to pertussis toxin is not an absolute requirement for prevention of infection with B. pertussis, evidence is provided that anti-toxin will eliminate toxic effects owing to secretion of pertussis toxin.

Allantois↗

Gastrulation in the sea urchin embryo: a model system for analyzing the morphogenesis of a monolayered epithelium.

Processes of gastrulation in the sea urchin embryo have been intensively studied to reveal the mechanisms involved in the invagination of a monolayered epithelium. It is widely accepted that the invagination proceeds in two steps (primary and secondary invagination) until the archenteron reaches the apical plate, and that the constituent cells of the resulting archenteron are exclusively derived from the veg2 tier of blastomeres formed at the 60-cell stage. However, recent studies have shown that the recruitment of the archenteron cells lasts as late as the late prism stage, and some descendants of veg1 blastomeres are also recruited into the archenteron. In this review, we first illustrate the current outline of sea urchin gastrulation. Second, several factors, such as cytoskeletons, cell contact and extracellular matrix, will be discussed in relation to the cellular and mechanical basis of gastrulation. Third, differences in the manner of gastrulation among sea urchin species will be described; in some species, the archenteron does not elongate stepwise but continuously. In those embryos, bottle cells are scarcely observed, and the archenteron cells are not rearranged during invagination unlike in typical sea urchins. Attention will be also paid to some other factors, such as the turgor pressure of blastocoele and the force generated by blastocoele wall. These factors, in spite of their significance, have been neglected in the analysis of sea urchin gastrulation. Lastly, we will discuss how behavior of pigment cells defines the manner of gastrulation, because pigment cells recently turned out to be the bottle cells that trigger the initial inward bending of the vegetal plate.

Animals↗

The mechanism of action of coculture on embryo development in the mouse model: direct embryo-to-cell contact and the removal of deleterious components.

OBJECTIVE: To elucidate the mechanism for the mode of action of coculture by the use of a coculture system for mouse one-cell embryos with human oviductal epithelial cells. DESIGN: Prospective, controlled in vitro experimental study. SETTING: Academic research laboratory. ANIMAL(S): Female ICR strain mice aged between 6 and 8 weeks. INTERVENTION(S): Flushed one-cell embryos were cultured in human tubal fluid medium alone (control), in coculture system with human oviductal cells, in five kinds of conditioned media, and in a contactless coculture system using a cell-culture insert. MAIN OUTCOME MEASURE(S): The percentage of the embryos developed to hatching blastocyst stage and the level of superoxide anion in the supernatant from each culture condition. RESULT(S): The rates of embryo development to the hatching blastocyst stage were significantly higher in the coculture group (43%) than in the control group (none) (P <.05). The embryo development rate in the control group was similar to that of the embryos in the five kinds of conditioned media. The effects of coculture on embryo development disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in the coculture group compared to the control group. CONCLUSION(S): The present coculture system overcomes the two-cell block in vitro and improves the embryo development. The beneficial effect may be a result of direct cell-to-cell contact between the embryo and helper cells and the removal of deleterious components from medium, rather than a result of embryotrophic factors.

Animals↗

Oxidized derivatives of 7-dehydrocholesterol induce growth retardation in cultured rat embryos: a model for antenatal growth retardation in the Smith-Lemli-Opitz syndrome.

7-Dehydrocholesterol accumulates in fetuses affected by the Smith-Lemli-Opitz syndrome as a result of a deficit in the ultimate step of cholesterol synthesis catalyzed by Delta7 reductase. Rat embryos explanted at gestation day 10 and cultured for 48 h in the presence of the Delta7 reductase inhibitor AY 9944 were used as a model to discriminate between the beneficial effect of supplementation with cholesterol and the deleterious effect of supplementation with 7-dehydrocholesterol. Cholesterol supplementation in the form of mixed cholesterol/lecithin liposomes added to serum serving as the culture medium restores the growth of embryos which is markedly decreased in the presence of the inhibitor. 7-Dehydrocholesterol under identical conditions does not restore growth and impairs the beneficial effect of cholesterol added simultaneously. UV-photooxidation of 7-dehydrocholesterol-supplemented culture medium enhances its embryotoxicity, which suggests uptake by the embryo of toxic by-products formed from 7-dehydrocholesterol. By contrast photooxidation of cholesterol-supplemented culture medium does not induce embryotoxicity. alpha-Tocopherol reduces the toxicity of photooxidized 7-dehydrocholesterol supplementing the culture medium. We conclude that 7-dehydrocholesterol does not fulfill the cholesterol requirement of the developing embryos and exerts an additional embryotoxic effect probably via oxidized by-products. This could explain the antenatal growth retardation of SLOS by a blockage of the maternal compensatory cholesterol influx.

Animals↗

Pharmacological inactivation of the endothelin type A receptor in the early chick embryo: a model of mispatterning of the branchial arch derivatives.

In the present study, we have applied an antagonist treatment to the chick embryo in ovo in order to demonstrate and dissect the essential roles of the endothelin type A (ETA) receptor in the embryonic development. We have cloned, sequenced and expressed the cDNA of the chick ETA receptor and shown that its affinity for endothelin antagonists is very similar to that shown by its mammalian counterparts. We have studied the spatio-temporal expression pattern of this receptor by in situ hybridization and shown that there is a high level of its mRNA within the mesenchyme of the branchial arches at E3-E5, in keeping with the direct effect of endothelin-1 (ET-1) on the fate of this region of the embryo. Unlike the endothelin type B (ETB) receptor mRNA, ETA mRNA is not expressed in neural crest cells during emigration from the neural tube, but is detected in neural crest-derived ectomesenchyme of the branchial arches. Finally, the functional involvement of this receptor in craniofacial and cardiovascular organogenesis was assessed by selectively inactivating the ETA receptor with specific antagonists applied during the time period corresponding to the expression of the ETA receptor and colonisation of the branchial arches. Embryos treated by these antagonists show a severe reduction and dysmorphogenesis of the hypobranchial skeleton, as well as heart and aortic arch derivative defects. This phenotype is very similar to that obtained in mice by gene inactivations of ET-1 and ETA. These results are observed with ETA antagonists but not with an ETB antagonist, and are dependent on the dose of the antagonists used and on the time of application to the embryo. Altogether, these data strongly show that the ET-1/ETA pathway, in chicken as in mammals, is a major factor involved directly and functionally in morphogenesis of the face and heart. This experimental model of pharmacological inactivation of a gene product described in this study offers a simple and rapid alternative to gene inactivation in mouse. This strategy can be applied to other ligand-receptor systems and extended to compounds of various chemical and functional natures.

Amino Acid Sequence↗

From oocyte to embryo: a model, deduced from in vitro fertilization, for natural selection against chromosome abnormalities.

A cytogenetical analysis was performed on 151 unfertilized oocytes, 22 fertilized eggs at the pronuclear stage, and 108 cleaved embryos obtained in the course of in vitro fertilization (IVF). Thirty-two per cent of unfertilized oocytes were abnormal, carrying nullisomies or disomies, mainly of D and G chromosomes, and a structural anomaly (Gq-) in one case. Fertilized eggs showed frequent asynchronism in the development of pronuclei and only 2 out of 8 karyotyped pronuclei were normal. Cleaved embryos were classified according to the number of pronuclei observed 17 hours after insemination. One per cent displayed a single pronucleus, and haploid chromosome complements were found in the corresponding cleaved embryos which were considered to be parthenotes. The rate of chromosome abnormalities of diploid eggs depended on their morphological aspect. Healthy cleaved embryos carried 12.5% of anomalies while this rate reached 37% in fragmented embryos (p less than 0.05). Lastly, 6% of fertilized eggs displayed three pronuclei or more. Only 41% of the corresponding embryos were triploid. Diploidy or diploidtriploid mosaicism were often encountered. This leads to a 21% rate of abnormalities in the preimplantation embryos. Parental karyotyping and HLA typing were carried out in a series of eight couples with in vitro idiopathic infertility or recurrent embryo degeneration in vitro. No abnormality was noted. According to these results, a model of natural selection of normal conceptuses is proposed.

Aneuploidy↗

Developmental toxicity research of ginsenoside Rb1 using a whole mouse embryo culture model.

BACKGROUND: Ginseng has been widely used around the world for many years. Knowledge is limited, however, on its effects on embryonic development. METHODS: Whole embryo culture was used to explore the developmental toxicity of ginsenoside Rb1 (GRb1) on mouse embryos. All embryos were exposed to different concentrations of GRb1, and scored for their growth and differentiation at the end of the 48-hr culture period. RESULTS: Total morphological score decreased significantly at the concentration of GRb1 of 30 microg/ml and was further reduced at 50 microg/ml. Yolk sac was affected at the lower concentration of 15 microg/ml. Developments of midbrain, forebrain, and optic system were relatively sensitive to GRb1 and were affected at the concentration of 30 microg/ml. Allantois, flexion, branchial arch, and limb buds were affected at 50 microg/ml. At this concentration, the embryonic crown-rump length, head length, and somite number were also reduced significantly compared to the control group. CONCLUSIONS: These results suggest that GRb1 has teratogenic effect during the mouse organogenetic period. We suggest that before more data in humans is available, ginseng should be used with caution by pregnant women in the first trimester.

Abnormalities, Drug-Induced↗

Embryo-dependent induction of uterine receptivity assessed by an in vitro model of implantation in mice.

To investigate the molecular basis of initial interactions between the trophoblast and the uterine epithelium at implantation, we developed an in vitro model for implantation in mice using primary uterine organ cultures as the substrate. In this model, embryos attached to the endometrial explants within 60 min. We were able to observe the attachment between embryos and endometrial explants in real time. Using this model, we investigated the attachment of embryos to uterine strips cultured from pregnant mice, pseudopregnant mice, pseudopregnant mice that had received hCG injections, and pregnant mice whose utero-tubal junctions had been ligated on Day 2 of pregnancy. The serum levels of progesterone in pregnant and pseudopregnant mice were also compared. The percentages of embryos that attached to uterine strips cultured from pregnant mice on Days 1-4 were low (0.0-15.0%). However, by Days 5-7, percentages of attachment increased markedly (66.8-74.8%). These levels were significantly higher than those observed for pseudopregnant mice (0.0-18.0%) and pseudopregnant mice that had received hCG injections (0.0-15.0%). The percentages of attachment to the ligated horns of pregnant Day 5 uteri (58.3-61.5%) were significantly higher than that of pseudopregnant mice (10.0%). The percentages of attachment to the ligated horns of pregnant Day 6 and 7 uteri decreased significantly (3.2%-19.4%). On the other hand, the percentages of attachment to the unligated horns of pregnant Day 6 and 7 mouse uteri did not decrease significantly (61.1-70.0%). The serum progesterone levels of pseudopregnant mice were not significantly lower than those of pregnant mice on Days 1-5.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Air fluid versus fluid-only models of embryo catheter loading: a systematic review and meta-analysis.

The objective of this systematic review was to determine the beneficial or detrimental effect of using air bubbles to bracket the embryo-containing medium during embryo transfer. To test this theory, a meta-analysis of randomized trials comparing air fluid versus fluid-only methods was performed. The primary outcome measures were live birth, ongoing and clinical pregnancy rates. The secondary outcome measures were the rates of implantation, miscarriage, multiple and ectopic pregnancies and retained embryos. Electronic (e.g.PubMed, EMBASE, Cochrane Library) and hand searches of the literature revealed two included studies (298 women). Meta-analysis was conducted using the Mantel-Haenszel method (fixed-effect model). For the primary outcome measures, there were no significant differences between the two methods with regards to live birth (OR = 1.34; 95% CI = 0.59-3.07), ongoing pregnancy (OR = 1.34; 95% CI = 0.59-3.07) and clinical pregnancy (OR = 1.13; 95% CI = 0.70-1.83) rates. For the secondary outcomes, there were no significant differences between the two groups. In conclusion there is insufficient evidence to suggest that the fluid-only method is superior to the use of air brackets during embryo loading. There is a need for well-designed and powered randomized trials to determine any possible benefit to either method.

Abortion, Spontaneous↗

Epigenetic Reprogramming and Zygotic Genome Activation in Human Preimplantation Development: Mechanisms, Models, and Translational Prospects.

PURPOSE: Early human embryogenesis unfolds through a tightly coupled sequence of events-clearance of maternal transcripts, remodeling of parental chromatin, zygotic genome activation (ZGA), lineage segregation, implantation, and post-implantation patterning-accompanied by epigenetic reprogramming, including X-chromosome dosage compensation around the time of implantation. This review aims to synthesize recent advances in understanding this developmental program and to consider their implications for reproductive medicine. METHODS: I review recent literature on human early embryogenesis, with particular emphasis on findings enabled by single-cell genomics and stem-cell-based embryo modeling, and integrate these insights to identify human-specific features of early development. RESULTS: These approaches have made previously inaccessible aspects of human early embryogenesis experimentally tractable, revealing molecular and epigenetic features that distinguish human development from that of model organisms, including species-specific dynamics of ZGA, maternal transcript clearance, chromatin reprogramming, and X-chromosome dosage compensation. CONCLUSIONS: Advances in single-cell genomics and embryo modeling are transforming our understanding of human early embryogenesis. Building on these insights, while recognizing their current limitations, I propose a vision for improving reproductive medicine, including the potential for next-generation embryo selection strategies.

Journal Article↗

Unveiling the establishment of left-right asymmetry in the chick embryo.

Vertebrates display striking left-right asymmetries in the placement of internal organs, which are concealed by a seemingly bilaterally symmetric body plan. The establishment of asymmetries about the left-right axis occurs early during embryo development and requires the concerted and sequential action of several epigenetic, genetic and cellular mechanisms. Experiments in the chick embryo model have contributed crucially to our current understanding of such mechanisms and are reviewed here. Particular emphasis is given to the elucidation of a genetic network that conveys left-right information from Hensen's node to the organ primordia, characterized to a significant degree of detail in the chick embryo. We also point out a number of early and late events in the determination of left-right asymmetries that are currently poorly understood and for whose study the chick embryo model presents several advantages. We anticipate that the availability of the chick genome sequence will be combined with multidisciplinary approaches from experimental embryology, biophysics, live-cell imaging, and mathematical modeling to boost up our knowledge of left-right organ asymmetry in the near future.

Animals↗

Possible lethal enhancement of toxins from putative periodontopathogens by nicotine: implications for periodontal disease.

AIM: To test the hypothesis that lethal synergy in the chick embryo model may occur between nicotine and bacterial products (cell-free extracellular toxins and cell lysates) of five putative periodontopathogens. METHODS: The lethality of cell-free extracellular toxins and cell lysates of five periodontal species was assessed with or without nicotine in the chick embryo assay system. Ten putative periodontopathogens (five species) were studied: Prevotella intermedia (n = 5), Porphyromonas gingivalis (n = 1), Porphyromonas asaccharolytica (n = 1), Fusobacterium nucleatum (n = 2), and Fusobacterium necrophorum (n = 1). RESULTS: Simultaneous testing of cell-free extracellular toxins from isolates W50, PS2, PS3, PS4, and PS5 and nicotine resulted in a percentage kill significantly greater than expected (Fisher's Exact test). Simultaneous testing of cell lysates from isolates W50, PS2, and PS5 and nicotine resulted in a percentage kill significantly greater than expected (Fisher's Exact test). CONCLUSIONS: Lethal synergy in the chick embryo model may occur between nicotine and toxins from putative periodontopathogens (both cell-free extracellular toxins and cell lysates). This may be an important mechanism by which smoking increases the severity of periodontal disease.

Animals↗

Fish and amphibian embryos--a model system for evaluating teratogenicity.

Developmental stages of fish and amphibians were used to evaluate reproductive impairments produced by environmental contaminants. Using static-renewal and flow-through procedures, exposure was initiated at, or soon after, fertilization and maintained through 4 days posthatching. Trace concentrations of many environmental toxicants produced high frequencies of mortality and teratogenesis. When developmental stages of the trout were exposed to inorganic mercury, 10% control-adjusted mortality was observed at 1 microgram/L and frequencies of terata in hatched populations ranged from 8% to 43% for exposure concentrations of 2 to 25 micrograms/L. In similar tests with the narrow-mouthed toad, 45% mortality occurred when mercury was administered at 1 microgram/L; frequencies of teratogenesis ranged from 8% to 17% at exposure concentrations of 1 to 5 micrograms/L; and complete mortality occurred at 7 micrograms/L. Boron-induced teratogenesis in trout ranged from 5% at 1 microgram/L to 51% at higher exposure levels. When catfish embryos were exposed to atrazine at 0.06, 4.8, and 46.7 mg/L, terata appeared in surviving populations at frequencies of 4%, 69%, and 100%, respectively. Fish and amphibian developmental stages constitute simple and effective models for 1) investigating mechanisms of teratogenesis, 2) evaluating the impact of environmental toxicants on aquatic biota, and 3) identifying environmental teratogens which may be of concern to human health.

Amphibians↗

Cytomegalovirus infection of the central nervous system stem cells from mouse embryo: a model for developmental brain disorders induced by cytomegalovirus.

Cytomegalovirus (CMV) is the most frequent infectious cause of developmental disorders of the central nervous system (CNS) in humans. Infection of the CNS stem cells seems to be primarily responsible for the generation of the brain abnormalities. In this study, we evaluated the infectivity of murine CMV (MCMV) in epidermal growth factor (EGF)-responsive CNS stem cells prepared from fetal mouse brains, and studied the effect of infection on growth and differentiation of the stem cells. The CNS stem cells were permissive for MCMV infection, although MCMV replication was slower than in mouse embryonic fibroblasts. MCMV infection inhibited the growth and DNA replication of the stem cells. A clonogenic assay revealed that MCMV infection suppressed generation of colonies from single stem cells. When uninfected stem cells were induced to differentiate, a decrease in expression of the primitive neuroepidermal marker nestin was observed by immunocytochemistry and flow cytometry, whereas expression of neurofilament and glial fibrillary acidic protein (GFAP) were induced. In virus-infected CNS stem cells, nestin expression was retained, whereas the expression of neurofilament was more severely inhibited than that of GFAP in these cells. Two-color flow cytometry showed that differentiated glial precursor cells were preferentially susceptible to MCMV infection. MCMV-infected and uninfected CNS stem cells were transplanted into the neonatal rat brains. The reduced number of infected stem cells were engulfed into the subventricular zone and expressed GFAP, but did not migrate further, in contrast to the uninfected stem cells. These results suggest that suppression of the growth of the CNS stem cells and inhibition of the neuronal differentiation by CMV infection may be primary causes of disorders of brain development in congenital CMV infection.

Animals↗