113 results about "Blastocyst" patented technology

The invention discloses an image detection and evaluation method for an unfrozen form of a fertilized egg embryo. The method comprises the following steps: acquiring a time sequence image sequence after the fertilized egg embryo is unfrozen; performing image standardization processing on the time sequence image sequence to generate a standardized embryo image set; extracting multi-dimensional embryo quality parameters from the standardized embryo image set; wherein the multi-dimensional embryo quality parameters comprise a morphological static parameter set, a dynamic change parameter set and a texture feature parameter set; generating a blastocyst formation rate based on the multi-dimensional embryo quality parameters; outputting a third-level embryo quality evaluation result according to a comparison relationship between the blastocyst formation rate and a preset threshold interval; wherein the three-stage embryo quality evaluation result comprises an excellent embryo grade, a qualified embryo grade and an abnormal embryo grade. The method has the following advantages and effects: full-automatic and multi-dimensional quantitative evaluation of the thawing process of the unfrozen embryos is realized, so that the scientificity of embryo screening and the success rate of clinical pregnancy are improved.

The invention belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a glucose slow-release structure used in an embryo in-vitro culture solution, a preparation process and a use method. The glucose slow-release structure comprises at least one inner layer containing glucose and a biodegradable material outer layer wrapping the inner layer, and the biodegradable material outer layer comprises a matrix with a plurality of micropores and at least one sealing material layer used for sealing pores of the micropores. According to the invention, gradient release from low sugar in the cleavage stage to high sugar in the blastocyst stage is realized, midway liquid change or artificial sugar supplement is not needed, the operation is obviously simplified, and pollution and mechanical stress risks are reduced.

Described are small molecule cocktails that enable de novo derivation and long-term culture of bovine extraembryonic endoderm cells (bXENs). Methods of using the small molecule cocktails to generate bXENs are also described. Methods of using the bXENs to form blastocyst models or to maintain the stemness of bovine ESCs and prevent them from differentiation are also described.

The invention relates to the technical field of livestock and poultry breeding, in particular to a goat hybridization breeding method capable of achieving complete-cycle nutrition enhancement. The core of the method is that systematic nutrition intervention runs through the whole breeding process. Before hybridization, performing differential nutrition pretreatment on the parents for 60 days: feeding the female parents with a breeding nutrition enhancer added with components such as D-chiro-inositol and N-acetylcysteine, and supplementing sperm motility enhancer containing acetylated L-carnitine and ergothioneine to the male parents. In the gamete treatment stage, the oocytes are subjected to in-vitro maturation culture in a special culture solution containing components such as D-chiro-inositol and a growth differentiation factor 9; after the sperms are subjected to gradient centrifugal screening, completing in-vitro fertilization in an optimized fertilization culture solution; after the embryos develop into blastocysts in the sequential culture system, carrying out laparoscopic transplantation, and carrying out staged precise nutrition culture on the pregnant ewes. According to the method, through multi-level nutrition enhancement, the reproductive capacity of the goats and the growth performance of offspring are effectively improved.

It is revealed that an organ such as pancreas can be regenerated by utilizing a fact that the deficiency of an organ is complemented by injecting an induced pluripotent stem cell (iPS cell) into a developed blastocyst in a blastocyst complementation method. Thus, the present invention has solved the above-described object. This provides a method for producing a target organ, using an iPS cell, in a living body of a non-human mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a different individual mammal that is an individual different from the non-human mammal.

Provided are a reagent combination or a kit for constructing an embryoid and the use thereof. The reagent combination or the kit can be used for inducing a stem cell to produce a blastocyst-like lineage precursor cell, a blastocyst-like cell and / or an embryoid, which are based on small-molecule induction and independent of transgenes, and have a single cell source. The produced blastocyst-like lineage precursor cell and blastocyst-like cell have a full blastocyst lineage, and are seed cells having balanced developmental competence and totipotency, which solves the problem of unbalanced cell maturation in the prior art. The produced embryoid is highly similar to natural embryos in terms of morphological characteristics and transcriptomic characteristics, has the capability of highly reproducing post-implantation embryonic development and / or gastrulation, which is beneficial for research on embryonic development in vitro. The method for inducing the stem cell to produce the blastocyst-like lineage precursor cell, the blastocyst-like cell and / or the embryoid by using the reagent combination or the kit is simple, and has high applicability and high embryoid construction efficiency.

The present invention relates to a medium composition for in vitro fertilization and / or in vitro culture comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a method using same. The compound or composition suppresses reactive oxygen species and lipid peroxidation in ovarian granulosa cells to preventing cytotoxicity and mitochondrial dysfunction caused by apoptosis and ferroptosis, leading to an improvement in the quality of aged oocytes, and to increase the developmental rate and blastocyst formation rate of pre-implantation embryos, thereby improving the efficiency of in vitro fertilization. In addition, treatment with the medium composition during the vitrification process of in vitro–fertilized embryos improves re-expansion and survival rates, thereby reducing structural and metabolic damage. Accordingly, the present invention can be advantageously applied to an assisted reproductive technology for treating infertility and subfertility, and in vitro fertilization and / or in vitro culture for improving the propagation efficiency of livestock.

The invention provides an oocyte in-vitro maturation culture solution additive and application thereof. The additive is tert-butylhydroquinone. By adding 100 [mu] g / mL of TBHQ into a basic culture solution, the first polar body discharge rate of oocytes is remarkably increased to 77.29%, the ROS level is reduced, and the blastocyst rate of in-vitro fertilized embryos is increased. The technology is suitable for breed conservation of livestock genetic resources such as sheep and can replace live livestock transportation to reduce the cost.

The present invention discloses a blastocyst expansion and puncture device, which includes: a base, a vertical shaft installed on the base, an expansion sac covered on the vertical shaft and capable of expansion, a plurality of mold flaps, a mold-closing mechanism for driving the mold flaps to open or close, puncture holes provided on the mold flaps, and a plurality of puncture components, each puncture component including a puncture mounting plate, a plurality of puncture needles installed on the puncture mounting plate and penetrated through the puncture holes, and a puncture drive part for driving the puncture mounting plate to move in a direction toward and out of the mold; the expansion molding is completed by limiting the shape of the blastocyst after expansion through the closed mold flaps, and when the expansion molding is completed, the puncture mechanism performs puncture to form an exhaust hole on the surface of the blastocyst, and two processes can be completed by one tooling, thereby greatly improving the processing efficiency of the blastocyst.

The invention relates to the technical field of construction of conditional gene knockout models, and discloses a construction method of a mouse Oct4 conditional gene knockout model based on quadruple sgRNA collaborative targeting, which comprises the following steps: step 1, designing four sgRNAs, namely sgRNA1, sgRNA2, sgRNA3 and sgRNA4, in which the sgRNA1 and the sgRNA2 target 5'end and 3 'end non-coding regions of a second exon of an Oct4 gene, and the sgRNA3 and the sgRNA4 target structural domain conserved sequences; step 2, preparing an electrotransfection solution, wherein the electrotransfection solution is composed of Cas9mRNA, the four sgRNAs and an electrotransfection buffer solution; step 3, carrying out electroporation treatment on the fertilized eggs of the mice; step 4, culturing the fertilized eggs subjected to the electrotransformation treatment to a blastocyst stage; and 5, carrying out genotype identification on the cultured blastocyst. According to the scheme, the sgRNA combination and embryo electrotransfection system is optimized, the mouse Oct4 conditional gene knockout model is efficiently and accurately constructed, a reliable tool is provided for regulation mechanism research of Oct4 in mouse early embryo trigerm differentiation, and the medical transformation value of Oct4 is excavated.

The invention discloses a line establishment method and a culture system of bovine trophoblast stem cells. The stem cell line is obtained by separation and induction from bovine blastocyst. The embryonic stem cell line has stability. The method can be applied to various life science and medical fields such as bovine animal breeding and breeding, gene editing models, animal cloned donor cells, medical musculoskeletal injury experiments, drug development carriers, vaccine production cell banks, embryo quality analysis and the like, and can be applied to large-scale production and application.

This invention discloses a method and system for embryo quality detection, relating to the field of embryo detection technology, including the following steps: S1: fertilized embryos are continuously cultured in an in vitro culture system containing specific metabolic marker detection components, and continuous dynamic imaging of the embryos is performed to obtain a complete dynamic developmental image sequence; S2: the dynamic developmental image sequence is segmented, and temporal developmental features are captured, identifying key time points such as pronucleus appearance and disappearance, cleavage, and blastocyst formation, and generating a segmentation mask corresponding to each frame of the dynamic developmental image sequence; this invention achieves full-process time-difference imaging to capture the temporal features of embryonic development, without trauma or interference, avoiding subjective bias in static observation; by segmenting the dynamic developmental image sequence and capturing temporal developmental features, the segmentation accuracy is high and the anti-interference ability is strong.

The present application discloses an artificial propagation-hybridization combined breeding method for meat goats, which relates to the technical field of animal breeding. The specific steps include: selecting a goat breed with good meat performance as the maternal parent and selecting a goat breed with a large body as the paternal parent; culturing the maternal oocytes in vitro to mature oocytes containing the first polar body, using a temperature gradient screening device to screen out highly motile sperm, fertilizing the mature oocytes with the highly motile sperm in vitro, culturing them to a hybrid blastocyst morphology, transplanting them into the maternal body to develop until natural delivery, and obtaining a hybrid F1 generation; backcrossing the F1 generation with the maternal parent, and backcrossing the backcross offspring with the maternal parent multiple times until the breeding goal is met, and then cross-crossing and fixing them to obtain a new breed / line with excellent traits; improving the sperm screening efficiency by utilizing the screening device, ensuring the quality of the screened sperm, and thus significantly improving the fertilization efficiency and embryo survival rate.

The application relates to a mouse model construction method for psoriasis research based on SBSN 600 lysine dihydroxyisobutyryl modification, which comprises the following steps: first step, constructing a plasmid comprising a DNA fragment of a SBSN gene sequence, wherein 600 sites of the SBSN sequence are mutated from lysine (K) to threonine (T); second step, culturing mouse embryonic stem cells; third step, electrically transforming the embryonic stem cells; fourth step, drug screening; fifth step, culturing the embryonic stem cells screened in the fourth step; sixth step, injecting the embryonic stem cells cultured in the fourth step into mouse blastocysts, then culturing the mouse blastocysts, and transplanting the cultured blastocysts into the uterus of a pseudopregnant female mouse, wherein the mouse delivered by the female mouse is a chimera mouse; and seventh step, breeding the chimera mouse and identifying the mouse genotype through PCR.

The present disclosure relates to a method for constructing a mouse model with short telomere. Specifically, the present disclosure obtains fertilized eggs by fertilizing the sperms and eggs of mice in vitro, and the fertilized eggs are cultured to the blastocyst stage in vitro and then transferred into the surrogate female mouse for development, thereby producing mice with short telomere. The method of the present disclosure does not require gene editing, has a short modeling cycle, and the effect is reliable and stable. By changing only the environment during embryo transfer, the telomere elongation process in the embryo is interfered, so as to successfully construct a progeny model with short telomere. There is no significant effect on the reproductive rate of female mice. Therefore, the present disclosure can provide a method for constructing a mouse model with short telomere for exploring the mechanism of telomere shortening and studying telomere-related phenotypes such as aging.

This invention relates to the fields of biomedical engineering and embryo quality assessment, and particularly to a biomarker for assessing the quality of IVM-derived embryos and its application. The biomarker includes time point parameters t2 and t9 of the IVM-derived embryos, and time interval parameters S2, t9–t7, t9–t8, or tSB–t9. A method for assessing the quality of IVM-derived embryos based on the aforementioned biomarker has been established. This method combines non-invasive temporal imaging technology with morphodynamic multi-parameter analysis, providing a reliable tool for assessing the blastocyst formation potential and chromosomal euploidy of IVM-derived embryos in clinical practice, thus potentially expanding the application of IVM technology in assisted reproductive technology.

This invention discloses a method, apparatus, electronic device, and storage medium for blastocyst quality assessment. The method includes: acquiring a sequence of images of a target blastocyst to be tested, the sequence comprising multiple images of the target blastocyst at different time points; processing the sequence of images of the target blastocyst to be tested based on a blastocyst quality assessment model to obtain a quality assessment result; the blastocyst quality assessment model includes a global feature extraction module, a temporal feature extraction module, and a result determination module; the global feature extraction module sequentially extracts global features from the images to be tested in the sequence, obtaining global features corresponding to each image; the temporal feature extraction module extracts temporal features from the global features corresponding to each image, obtaining temporal features; and the result determination module classifies the temporal features to obtain a quality assessment result, achieving accurate assessment of blastocyst quality and providing accurate data support for subsequent decision-making.

The invention belongs to the technical field of biology, and particularly relates to application of astragaloside in regulation of parthenogenetic activation embryo development. According to the method disclosed by the invention, the maturation rate of the oocytes can be obviously improved by adding substances such as astragaloside into the sheep oocyte in-vitro maturation liquid. The preparation method comprises the following steps: firstly, respectively adding lycopene, astragaloside, isoliquiritigenin, forsythin and baicalein into an oocyte maturation solution, and screening out the optimal dosage of each additive according to the first polar body discharge rate, the cleavage rate and the blastocyst rate; and finally, adding an additive combination into the oocyte maturation liquid according to the screened optimal dosage, detecting the first polar body discharge rate, the cleavage rate and the blastocyst rate to determine an optimal composition, and detecting the levels of ROS, GSH, mitochondrial membrane potential, oxidative stress and apoptosis related genes of the optimal composition to further determine the application effect of the composition.

The application provides a culture medium and method for stem cell differentiation into macrophages, and relates to the technical field of biotechnology.The culture medium provided by the application comprises six kinds of culture media, the components of which are clear, and the culture media and cytokines are prepared without animal sources, so that exogenous substances are avoided, the differentiated cells are safer, and clinical transformation can be seamlessly connected.The preparation of macrophages by using the culture medium does not need to pass through an EB process, so that the steps are simplified, the differentiation time is shortened, the purity, uniformity and stability of differentiation are improved.The method for differentiating macrophages provided by the application skips the complicated blastocyst formation stage, simplifies the differentiation steps, greatly shortens the differentiation period, reduces unknown risks, improves the differentiation efficiency and stability, can obtain a large amount of high-purity macrophages with killing capacity in a short period, and also provides the possibility of full-closed automatic production for large-scale production preparation, and solves the bottleneck of industrialization difficulty.

The present disclosure relates to a method implemented by a computer system, and an oocyte quality analysis system and a non-transitory computer-readable medium that stores instructions, and particularly relates to a system and method for assessing the viability of an oocyte. In some embodiments, an image sequence related to an oocyte drawn into a pressure tool is acquired. By using a segmentation model, objects related to the oocyte can be identified. On the basis of the identified objects, features such as morphological features and the drawing depth related to the oocyte can be determined. Then, at least some of the features can be input into a machine learning model to generate an oocyte grade that indicates the probability of the oocyte developing into a usable blastocyst. Optionally, the oocyte grade can be presented to a user by means of an interactive user interface.

The invention discloses an application of an Odf4 gene or protein as a biomarker in evaluating the ability of sperms to bind to oocytes. According to the invention, the interaction between Odf4 protein and sperm-egg binding key factors ZP3R and ADAM2 is found for the first time through experiments, and a sperm-egg recognition complex is formed; odf4 gene deletion can lead to significant reduction of the ability of sperms to bind to oocytes, and the formation rates of 2-cells, 4-cells, 8-cells and blastocysts of fertilized embryos are significantly reduced. On the basis, by detecting the expression level or functional integrity of the ODF4 in the sperms, the binding capacity of the sperms and oocytes and the development potential of fertilized embryos can be objectively evaluated. According to the invention, the limitation of low detection rate of recessive sperm-egg binding defects in the prior art is overcome, and an efficient molecular tool is provided for etiological typing, assisted reproduction scheme selection and fertility prediction of male infertility.

This invention provides an application and method of lysophosphatidylcholine (LPC) in promoting early development of reconstructed embryos, belonging to the field of embryo engineering technology. The invention first prepares reconstructed embryos, then adds lysophosphatidylcholine to the culture system for culturing. Lysophosphatidylcholine effectively promotes early development of the reconstructed embryos; at an optimal concentration of 5 μM, the blastocyst development rate increases to 24.56%, providing a novel and operable biochemical intervention target to solve the problem of low reconstructed embryo culture efficiency. This invention not only fills the gap in our understanding of the mechanism of action of LPC in this field, but also achieves controllability and reproducibility of the technical solution through precise definition of the optimal concentration. It provides key parameters for establishing standardized culture procedures and provides a more solid foundation for downstream applications such as developmental biology research and disease model construction that rely on high-quality reconstructed embryos, significantly accelerating the process of related scientific discovery and application development.

The invention relates to a low-temperature preservation method for sheep semen, and belongs to the technical field of animal reproduction. The invention provides a method for low-temperature preservation of sheep semen, which comprises the following steps: (1) collecting breeding male sheep semen by a false vagina method to obtain the collected sheep semen; (2) mixing the sheep semen collected in the step (1) with a semen diluent and fullerene to obtain a sheep semen diluent; and (3) preserving the sheep semen diluent under the condition of 2-8 DEG C. The sheep semen is preserved according to the method, the effective application time of sperm preservation can be remarkably prolonged, the blastocyst forming rate of in-vitro fertilization embryos can be remarkably increased, and the method is applied to reproduction technologies such as artificial insemination (AI) and in-vitro fertilization (IVF) of sheep, the utilization rate of high-quality breeding rams can be increased, and genetic resource protection and efficient breeding can be promoted.

The present invention discloses an ICSI operation method for improving the efficiency of producing sexed OPU embryos in vitro in dairy cows: frozen sexed sperm is treated using the BO flotation method, and the floated sperm is centrifuged and diluted with IVF. The treated sexed sperm is injected into OPU donor oocytes, activated twice with 5nM ionomycin and once with 2nM 6-DMAP, and then embryo culture is performed. The present invention significantly improves the blastocyst rate of embryos produced after ICSI eggs are activated, and can be used to stably produce high-quality sexed OPU embryos in dairy cows.

The invention provides application of ergothioneine in improving fertility of high-fertility-age female individuals, and belongs to the technical field of natural medicines. The invention provides an application of EGT in preparation of a product for improving fertility of a high-fertility-age female individual. In the embodiment of the invention, a high-fertility-age ICR female mouse is taken as an animal model, the influence of the EGT on the fertility of the high-fertility-age female mouse is verified from in-vivo and in-vitro aspects, and the result shows that the EGT has the effect of improving the in-vitro fertilization rate and blastocyst rate of oocytes of the high-fertility-age female mouse; the superovulation number and the litter number of high-childbearing-age female mice are increased. It is proved that the EGT has the effect of improving the fertility of high-fertility-age female individuals.

The application discloses a construction method of a spontaneous psoriasis mouse model and application thereof, and inserts a loxP site into an embryonic stem cell clone in a fatty acid synthase gene Fasn Chimeric mice are obtained through blastocyst injection, and through mating, a Fasn flox / flox Mouse; and a tool mouse Krt14-CreERT2 + / ‑ The tool mouse is mated with the tool mouse, and a mouse with a genotype of Krt14-CreERT2 + / ‑ ; Fasn flox / flox is screened, and the mouse model is obtained through continuous induction of tamoxifen. The spontaneous psoriasis mouse model constructed by the application is superior to an acute model which needs external induction, is convenient for long-term observation and intervention experiments, and provides a more ideal tool for researching a pathogenesis of psoriasis, screening and evaluating new drug candidate compounds for psoriasis.

The present discloses provides the use of tributyrin as an additive for embryonic development culture medium in vitro, in the present disclosure, the tributyrin is applied as an additive to the development culture medium of mouse embryos in vitro for the first time, the tributyrin can significantly increase the rate of blastocyst, and reduce the ROS content in the embryos, improve the mitochondrial membrane potential in the embryos, increase the ATP level and the expression of antioxidant genes in the embryos, improve the DNA methylation and histone modification level in the embryos, promote the embryos development in vitro; In addition, tributyrin, as natural antioxidant and apparent drug, which is safe, non-toxic and side effects; the tributyrin provides strong support for the efficient embryos development of human assisted reproductive technology, mammalian fertilization embryos, parthenogenetic embryos and somatic cell cloned embryos and other embryo engineering technologies in vitro.

The invention provides application of endothelin in oocyte in-vitro maturation and embryo in-vitro culture. New application of endothelin is found, the endothelin can improve the polar body discharge rate of oocytes matured in vitro, promote cytoplasm maturing and improve the normal arrangement proportion of organelles after maturing, certain promotion is provided for improvement of oocyte fertilization, and meanwhile subsequent blastocyst development is promoted. The invention further provides an in-vitro maturation culture solution of the sheep oocytes and an in-vitro maturation method of the sheep oocytes, the in-vitro maturation culture solution contains endothelin, and after the sheep oocytes are treated with the in-vitro maturation culture solution containing the endothelin concentration, the mitochondrial membrane potential level of the oocytes can be remarkably improved, and the survival rate of the sheep oocytes is increased. ROS (reactive oxygen species) in the oocytes is remarkably reduced, and the ATP (adenosine triphosphate) and GSH (glutathione) levels of the oocytes and the normal proportion level of spindles are improved; the maturation rate, the fertilization efficiency, the blastocyst number and the blastocyst rate are remarkably increased, in-vitro maturation of sheep oocytes and in-vitro development of early embryos are promoted, and the production efficiency of the sheep in-vitro embryos is effectively improved.

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing an embryoid and application of the reagent combination or kit. The reagent combination or the kit can be used for inducing stem cells to generate blastocyst-like pedigree precursor cells, blastocyst-like cells and / or embryoid-like cells, is based on small molecule induction, does not depend on transgenosis, and is single in cell source, and the generated blastocyst-like pedigree precursor cells and blastocyst-like cells have a whole blastocyst pedigree, and can be used for inducing stem cells to generate blastocyst-like pedigree precursor cells and blastocyst-like cells. The seed cells are balanced in developmental ability and have totipotency, and the problem of unbalanced cell maturation generated in the prior art is solved; the generated embryoid is highly similar to a natural embryo in morphological characteristics and transcriptome characteristics, has the capability of highly reproducing embryonic development and / or protointestinal movement after implantation, and is beneficial to in-vitro research of embryonic development; the reagent combination or the kit is used for inducing stem cells to generate blastocyst-like pedigree precursor cells, blastocyst-like cells and / or embryoid-like cells, and the method is simple, high in applicability and high in embryoid-like construction efficiency.