87 results about "Blastocytes" patented technology

The invention discloses a bovine oocyte in-vitro maturation culture solution as well as a preparation method and application thereof, and belongs to the technical field of animal reproduction and embryo engineering. The preparation method of the bovine oocyte in-vitro maturation culture solution comprises the step of adding mitoquinine mesylate into a conventional IVM culture solution, wherein the adding concentration of the mitoquinine mesylate is preferably 50 nmol / L. The bovine oocyte in-vitro maturation culture solution provided by the invention can effectively reduce the oxidative stress reaction of the oocytes in the in-vitro culture process, enhance the mitochondrial function and remarkably improve the bovine oocyte in-vitro maturation quality and the embryonic development potential, so that the blastocyst rate is increased, and the problem that the development potential of the existing oocyte IVM culture solution is insufficient is solved.

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.

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 belongs to the technical field of bioengineering, and particularly relates to application of RPS26 recombinant protein in promoting early embryonic development of animals. Experiments show that after the RPS26 recombinant protein is added in the early-stage in-vitro embryo culture process of the lamb goats, the blastocyst rate of the lamb goats can be increased to 40.0%; meanwhile, the cell proportion of cell mass in the blastocyst can be remarkably increased, the cell proportion of trophoderm is reduced, and the total number of blastocyst cells is not influenced, so that the blastocyst development rate can be remarkably increased and the blastocyst quality can be improved by utilizing the RPS26 recombinant protein.

The embodiment of the invention discloses an in-vitro fertilization embryo culture blastocyst formation rate prediction method and system. The method comprises the following steps: acquiring parameter information of an in-vitro fertilization embryo to be evaluated; the parameter information serves as input and is input into a pre-trained prediction model, a formation rate prediction result of the to-be-evaluated in-vitro fertilized embryo is obtained, and the pre-trained prediction model is obtained through training based on a LightGBM model; and if the formation rate prediction result of the to-be-evaluated in-vitro fertilized embryo is greater than a preset threshold value, using the to-be-evaluated in-vitro fertilized embryo for subsequent transplantation.

The present invention belongs to the field of molecular biology and genetics, and in particular relates to a method for editing a bovine gene based on Pro-iCHI. The present invention provides a method for editing a bovine gene based on Pro-iCHI. Protamine is transiently expressed in gene-edited b-haSCs, which are then injected to mature oocytes to obtain reconstructed embryos. Protamine can eliminate abnormal DNA methylation resulting from oocyte intracytoplasmic haSCs injection and enable the nucli to compress into sperm-like structures, and the obtained bovine Pro-iCHI embryos can successfully develop into blastocysts, with a blastocyst rate comparable to that of the embryos obtained by in vitro fertilization. Moreover, in the present invention, a protamine-encoding gene is inserted into a Saccharomyces cerevisiae protein expression vector for transient expression, which ensures that abnormal DNA methylation is erased, without integration into the genome resulting in the insertion of exogenous genes.

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.

A method of deriving and maintaining Salmonidae stem cells is provided, the method comprising deriving Salmonidae cells at the eyed or blastula stages of salmonid or salmonoid ova development, so as to produce cells with use in the food industry.

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 field of bioengineering technology, and in particular to a method for simultaneously collecting the inner cell mass and trophectoderm of mammalian blastocysts. The method includes the following steps: (1) after removing the zona pellucida of the blastocyst, the blastocyst is cut into an inner cell mass and a trophectoderm portion, and the cut trophectoderm portion is transferred to a culture medium for collection; (2) the inner cell mass portion obtained in step (1) is sequentially placed in an antibody treatment solution and a complement treatment solution for incubation; (3) the inner cell mass portion treated in step (2) is pipetted to obtain a purified inner cell mass. This method achieves simultaneous and efficient separation of two key cell lines from the blastocyst, fundamentally changing the traditional method's "resource waste" model of only obtaining the inner cell mass while discarding the trophectoderm, and providing more comprehensive experimental materials for embryonic development, placental biology, and stem cell research.

The present application relates to the field of animal embryo engineering technology, and provides a method for improving the development ability of bovine IVP embryos. In the production of bovine in vitro embryos, by adding PAPPA, ergothioneine and selenocysteine to the bovine oocyte in vitro maturation culture solution, early embryo culture solution, late embryo culture solution, freezing solution and thawing solution, the in vitro maturation performance of oocytes and the development performance and vitality of the subsequent obtained embryos can be effectively improved. Studies have shown that after adding the above additives, the maturation rate of oocytes, the cleavage rate, the blastocyst rate of embryos, and the freezing survival rate, the hatching rate and the ATP content of embryos after freezing preservation are significantly improved, which has important application value.

The invention relates to the technical field of artificial intelligence in assisted reproduction, in particular to an embryo optimization model construction method, device and equipment and a storage medium. Comprising the following steps: acquiring video data of embryonic development from a plurality of fertilized eggs to a blastocyst stage based on a delayed photography embryo culture technology; constructing time sequence image data according to the video data; the time sequence image data comprises multiple frames of continuous images; performing feature extraction on each frame of image of the time sequence image to obtain a high-dimensional feature corresponding to each frame of image; determining a comparison difference feature with the corresponding reference sample feature, performing fusion, and constructing a fusion feature corresponding to each frame of image; constructing a feature sequence of the target embryo development according to the fusion features; and acquiring a trained embryo quality grading model according to a time dependency relationship of the feature sequence for capturing embryo development. The model provided by the invention can optimize the utilization rate of embryos, reduce unnecessary repeated ovulation promoting cycles, and promote the development of reproductive medicine towards the precision direction.

The application provides a blastocyst three-dimensional fitting reconstruction method and device, electronic equipment and storage medium. The method comprises the following steps: acquiring a plurality of focal plane images of a blastocyst; identifying a trophoblast cell mask and an inner cell mass mask in the blastocyst based on the plurality of focal plane images; constructing a trophoblast cell grid model and an inner cell mass grid model according to the trophoblast cell mask and the inner cell mass mask; constructing a depth map of the blastocyst according to the plurality of focal plane images; performing image fusion on the plurality of focal plane images to obtain a texture image; performing plane mapping according to the texture image and the depth map to generate an internal three-dimensional point cloud model of the blastocyst; constructing a three-dimensional geometric model of the blastocyst according to the trophoblast cell grid model, the inner cell mass grid model and the internal three-dimensional point cloud model; and mapping the texture image to the surface of the three-dimensional geometric model to generate a blastocyst three-dimensional model. The reconstruction based on the method does not affect the development of the embryo, and is low in cost and safe and reliable.

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 provides a method for the cryopreservation and reconstitution of porcine embryos. During the in vitro maturation of porcine oocytes, porcine IVF embryo culture, and embryo freezing and thawing processes, the addition of TSA, 5-Aza-dC, Etomoxir, GSK-J4, and NR, either alone or in combination, improves the development and freezing efficiency of porcine IVF embryos. Experiments show that the oocyte maturation rate, embryo cleavage rate, and blastocyst rate of the groups adding TSA, 5-Aza-dC, and Etomoxir to the porcine oocyte in vitro maturation culture medium and porcine IVF embryo culture medium are significantly higher than those of the control group. Furthermore, the hatching rate of porcine IVF embryos after freezing and thawing is significantly higher in the groups adding GSK-J4 and NR to the freezing and thawing solutions than in the control group. This invention lays the foundation for the production of porcine in vitro fertilized embryos and related scientific research, providing an effective, safe, and feasible strategy for improving the production efficiency and quality of porcine in vitro embryos.

The invention discloses a culture solution for improving the development rate of in-vitro cultured pig blastocysts as well as application and a use method of the culture solution, and belongs to the technical field of livestock embryo engineering. The invention discloses a culture solution for improving the development rate of in-vitro cultured pig blastocysts, which is characterized in that a PZM3 culture solution is used as a basic culture solution, and 5-25ng / mL of tumor necrosis factor superfamily cell factor TNFSF12 is added. According to the present invention, the pig embryo needs to be subjected to in-vitro culture for 4 days in the TNFSF12-free basic culture solution, and then is transferred to the TNFSF12-containing basic culture solution so as to be continuously cultured for 2 days, such that the generation efficiency of the pig blastocyst can be effectively improved, and the support is provided for the application of the pig in-vitro fertilization, the somatic cell nuclear transfer, the embryo genome selection and other embryo engineering technologies in the agricultural breeding and the human medicine;

The invention discloses a culture solution for improving in-vitro culture blastocyst rate of bovine oocytes and application of the culture solution, and belongs to the technical field of cell in-vitro culture. According to the culture solution, kaempferol is added into a basic culture solution, and the addition concentration of the kaempferol is 0.01-1 mu M / L; preferably, the addition concentration of the kaempferol is 0.1 mu M / L. By optimizing the concentration of the kaempferol and the synergistic proportion of the kaempferol and basic nutritional ingredients, the action mechanism of the kaempferol for regulating and controlling the mitochondrial function of the oocyte and meiosis spindle assembly is defined, and the direct association of'oxidation resistance-development potential improvement 'is established, so that the blastocyst formation rate and embryo quality of in-vitro culture of the oocyte are remarkably improved, and the development potential of the oocyte is improved. The requirements of the fields of assisted reproduction, embryo engineering and the like on an efficient and stable oocyte in-vitro culture technology are met.

This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.

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 invention discloses a culture solution for promoting in-vitro maturation of bovine oocytes and application of the culture solution, and belongs to the technical field of assisted reproduction. The invention provides the culture solution which is composed of brand new components and is used for promoting in-vitro maturation of the bovine oocytes, support is provided for maturation of the oocytes from multiple aspects of carrier transportation, energy metabolism regulation, signal transduction, extracellular environment and the like, the in-vitro maturation quality of the bovine oocytes can be effectively promoted, and the survival rate of the bovine oocytes is increased. The quality and consistency of nucleus maturation and cytoplasm maturation of oocytes are obviously improved; the cultured mature oocytes are used for in-vitro fertilization, so that the fertilization proportion and the blastocyst hatching rate can be remarkably improved; the provided culture solution is appropriate in component number, clear in proportion, simple and easy to operate in preparation method, short in time consumption, good in repeatability and suitable for batch production, and has a good application prospect in maturation culture of bovine in-vitro oocytes.

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.

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 relates to the technical field of gene editing, in particular to a composition and a culture solution for promoting development of bovine gene editing cloned embryos and application of the composition and the culture solution. The composition (DLL3, Vercircon and ML604086) is added into an oocyte in-vitro maturation culture medium, and the DLL3, the Vercircon and the ML604086 cooperate with one another, so that apoptosis and autophagy injury of bovine oocytes are effectively reduced, the cleavage rate and blastocyst rate of embryos are increased, and the method has important practical value for improving the production efficiency and quality of gene editing cloned embryos.

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.