71 results about "Blastocyst" patented technology

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 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.

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.

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 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 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.

This disclosure relates to a system and method for generally evaluating the viability of oocytes. In some embodiments, an image sequence is acquired relating to oocytes aspirated into a pressure tool. Based on the image sequence and the pressure applied to the oocytes, morphological and mechanical features relating to the oocytes can be derived. At least some of these features can then be input into a machine learning model to determine metrics indicating the quality of the oocytes, where certain metrics may indicate blastocyst formation. Optionally, the determined oocyte quality information can be presented to the user via an interactive user interface.

The invention discloses application of a compound shown in a formula I in promoting in-vitro cultured oocyte development or in-vitro embryo development, and in the formula I, R1 is selected from halogenated alkyl; r2 is selected from H, halogen or C1-6 alkyl; n is selected from 0, 1, 2 or 3. According to the application, the expansion area of cumulus cells in the cumulus-oocyte complex can be increased; the oxidative damage of the cumulus-oocyte complex is relieved; the first polar body discharge rate of the oocytes is improved; the abnormal distribution rate of mitochondria in oocytes is reduced; the membrane potential of mitochondria in the oocyte is improved; and the cleavage rate and blastocyst formation rate of fertilized eggs can be improved, and the apoptosis rate of oocytes can be reduced.

This application relates to a method, apparatus, computer equipment, storage medium, and program product for the quantitative evaluation of trophectoderm. The method includes: acquiring target parameters such as the geometric parameters of the closed region containing the trophectoderm in the embryo to be evaluated, the total number of cells included, the cell area of ​​each cell, and the total number of adjacent cell pairs; obtaining standardized quantitative indicators, density indicators, and homogeneity indicators of the trophectoderm based on the target parameters; and obtaining evaluation results by quantitatively evaluating the trophectoderm based on the standardized quantitative indicators, density indicators, and homogeneity indicators. This application provides a calculable, standardized, and comparable quantitative evaluation method for trophectoderm, improving the objectivity, consistency, and repeatability of quantitative evaluation of trophectoderm in blastocysts.

An apparatus configured to facilitate delivery of an embryo to an implantation site on an endometrium of a patient is provided. The apparatus comprises one or more walls defining a cavity, the walls comprising a rim configured for adhering to the endometrium and defining an opening to the cavity. The walls comprise one or more throughgoing apertures, each sized to allow passage therethrough of intrauterine environmental molecules, and to prevent passage therethrough of a blastocyst. The material of the walls is configured to degrade in the intrauterine environment within a predetermined range of time.

Disclosed herein are chimeric blastocysts, such as chimeric blastocysts derived from a host blastocyst from a first mammalian species and having donor pluripotent stem cells from a second mammalian species, wherein the donor pluripotent stem cells have reduced expression or reduced biological activity of one or more proteins in the TLR / NF-kB signaling pathway or the p53 pathway. Methods of preparing chimeric blastocysts and methods of obtaining mammalian organs and tissues are also provided.

This invention provides a method for single-cell reduced-representation bifulfite sequencing (scRRBS) of embryo culture medium. Using medical waste (blastocyst culture medium) from in vitro fertilization (IVF) procedures as raw material, this invention allows for simultaneous dual analysis of embryonic chromosomal aneuploidy and DNA methylation status. It assesses embryonic developmental potential from a novel perspective, considering epigenetics and the embryo's response to its culture environment, providing a new reference for selecting the "right" embryos in assisted reproduction and strongly supporting improvements in the success rate of IVF cycles.

The present application belongs to the field of assisted reproductive technology, and particularly relates to a glucose slow-release structure for embryo in-vitro culture medium, a preparation process and a use method. The glucose slow-release structure comprises at least one glucose-containing inner layer and a biodegradable material outer layer covering the inner layer, wherein the biodegradable material outer layer comprises a substrate with a plurality of micropores and at least one sealing material layer for sealing the micropores. The present application realizes gradient release from low sugar in the cleavage stage to high sugar in the blastocyst stage, without the need for medium replacement or manual sugar supplementation in the middle, significantly simplifying the operation and reducing the risk of pollution and mechanical stress.

The invention belongs to the technical field of animal embryo engineering, and particularly relates to a cattle blastocyst development potential evaluation method and system based on unmarked imaging and lipid metabolism characteristics. The method comprises the following steps: performing laser unmarked multi-modal imaging on a cattle blastocyst to obtain a multi-modal fusion image including cattle blastocyst morphology, metabonomics and matrix structuring; quantitatively extracting morphological kinetic parameters, metabolic activity parameters and matrix structure parameters from the multi-modal fusion image, and constructing a multi-dimensional feature set; and training an evaluation model constructed based on a machine learning algorithm by adopting the multi-dimensional feature set, and obtaining a development potential score of the cattle blastocyst to be evaluated based on the trained evaluation model. According to the method, high-precision, non-invasive and objective prediction of the blastocyst in-vivo development outcome before transplantation is realized.

The invention provides a composition, a cell culture medium and application of the cell culture medium in establishment of trophoblast stem cells in bovine hatching blastocysts, and belongs to the technical field of cell engineering. The invention provides a composition. The composition comprises activin A, CHIR99021 and a recombinant human leukemia inhibition factor. After the composition is adopted to culture hatched blastocysts, bovine trophoblast stem cells are obtained in vitro, trophoblast cell specific markers are highly expressed in mRNA and protein levels, and the bovine trophoblast stem cells can be stably cultured in vitro for a long time. The composition not only can further enrich the culture system of the bovine trophoblast stem cells, but also can expand understanding of the difference of signal pathways causing the blastocysts in different stages of the cattle.

The utility model provides a capsule expansion drum device for air spring blastocyst pricking, which consists of a plurality of expansion drum bodies, the plurality of expansion drum bodies are synchronously expanded through a ventilation device, the outermost expansion drum body is connected with a main machine through a mounting shaft, and two adjacent expansion drum bodies are connected through a connecting shaft. The connecting shafts comprise the first connecting shaft and the second connecting shaft, the first connecting shaft is coaxially and fixedly installed on the expansion drum body on the front side, the second connecting shaft is coaxially and fixedly installed on the expansion drum body on the rear side, and the second connecting shaft is inserted into the first connecting shaft to slide and is fixed through a fixing part. The distance between the two expansion drum bodies is adjusted by adjusting the insertion depth between the first connecting shaft and the second connecting shaft, the requirement for fixing air spring blastocysts of different length specifications can be met, meanwhile, synchronous expansion of the multiple expansion drums can be achieved under the action of the ventilation device, and the requirement for fixing the blastocysts is met.

The present application relates to the assisted evaluation method of the second biopsy in the pre-implantation genetic testing. The blastocyst first biopsy process collects and cryopreserves the corresponding biopsy droplet; for the blastocyst with CNV detection failure or suspected abnormal CNV detection result in the first biopsy cell, the first biopsy droplet is thawed to complete genome amplification, CNV library construction, high-throughput sequencing and CNV analysis, and the second biopsy suggestion is given according to the detection result. If the biopsy droplet detects the abnormal aneuploidy or chimera clinically interpreted, it is suggested that the corresponding blastocyst has a high probability of chromosomal abnormality, and the second biopsy is not recommended; if the biopsy droplet does not detect the abnormal aneuploidy or chimera clinically interpreted, it is suggested that the corresponding blastocyst has a high probability of chromosomal normality, and the second biopsy is recommended. The present application proves that the first biopsy droplet contains detectable genetic material, and the consistency of the chromosomal ploidy detection result and the biopsy cell is more than 80%, which can assist the second biopsy decision and improve the decision efficiency and scientificity of PGT detection.