43 results about "Bovine embryo" patented technology

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The invention relates to the field of computer vision, in particular to a bovine embryo quality evaluation method and system based on a time sequence microscopic image, and the method comprises the steps: collecting a microscopic image and a timestamp, carrying out the embryo segmentation and inter-frame registration of the image, and obtaining the frame quality weight, morphological characteristics, image block embedding and topological data; respectively calculating a topological distance and an optimal transmission distance, determining a change point generation event sequence in combination with a frame quality weight, and aligning and outputting a penalty term based on a time Petri network; inferring a stage sequence and a quality posteriori based on the event sequence and a penalty term, and constructing a multi-instance set based on an event time window to obtain a multi-instance learning score; and fusing quality posteriori and multi-instance learning score to obtain an evaluation result, calculating a conflict coefficient, and relocating an event time window when a threshold value is exceeded to update the evaluation result. According to the method, rating consistency, noise immunity and traceability are improved.

The present disclosure relates to methods, products, and compositions useful for deriving bovine induced blastoids. Methods are described that allow for juvenile stem cell aggregation and induced blastocyst formation. The invention further provides an aggregation culture medium and an induced blastocyst-like culture medium which are suitable for promoting aggregation of the young stem cells and preparing the induced blastocyst-like. The bovine induced blastoids can be used in selection and breeding procedures, including the replication of pre-implant embryos with desired characteristics and in vitro breeding including a cross-generation breeding cycle from parent germ cells and / or embryos to progeny embryos.

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 discloses a kit for predicting the development state of a bovine embryo and a CNVR combination used by the kit. The kit comprises a product for detecting the expression quantity and / or copy number of the CNVR combination in the bovine embryo genome; the CNVR combination comprises CNVR-Vst5, CNVR-Vst11, CNVR-Vst7 and / or CNVR-Vst4, and the CNVR-Vst5, the CNVR-Vst11, the CNVR-Vst7 and the CNVR-Vst4 If the expression quantities and / or copy numbers of CNVR-Vst5, CNVR-Vst11, CNVR-Vst7 and / or CNVR-Vst4 in a bovine embryo genome subjected to in-vitro fertilization culture are obviously different from those of a normal bovine embryo, the development of the bovine embryo is predicted to be stagnated, and embryo implantation is not suggested. The method can be used for scientifically judging the transplantation strategy and decision of the bovine embryo, and has important application value.

The invention belongs to the technical field of cell biology, and particularly relates to a feeder-layer-free and serum-free bovine embryonic stem cell culture medium, a culture system and a culture method. The bovine embryonic stem cell culture medium comprises a conditioned culture medium and additives, the additives are IWR-1 and activin A, and the concentrations of the IWR-1 and the activin A in the conditioned culture medium are 2.5-5 [mu] M and 20-25 ng / mL respectively; the culture system comprises a bovine embryonic stem cell culture medium, gelatin with the mass concentration of 0.1-0.2% and a subculture medium, and the gelatin with the mass concentration of 0.1-0.2% is used for providing an attachment matrix for bovine embryonic stem cells; the subculture medium is prepared by adding Y27632 into a bovine embryonic stem cell culture medium. The cost of the extracellular matrix is reduced, and the feeder-layer-free bovine embryonic stem cell has high single cell passage efficiency and is convenient for gene editing operation.

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.

This invention belongs to the field of organoid culture technology, specifically involving BCL2 Application of genes in 3D cluster culture of stem cell-derived organoids. Starting on day 4 of bovine embryonic stem cell cluster culture, the cell cluster edges became indistinct, cells began to die and darken, and the cell clusters began to diffuse. By day 7, the cell cluster survival rate was only 14.2%. This invention utilizes the overexpression of genes in bovine embryonic stem cells... BCL2 Following gene expression, the cells were cultured in clusters using the same method. The overexpressed cell clusters showed clear edges, and their volume increased continuously with the number of culture days. Even on day 7, clear cell cluster edges were still observable, and no diffuse cell death was observed; the survival rate remained high at 85.54%. BCL2 Genes can improve the survival rate of early cultured cell clusters of organoids, which is of great significance for the normal morphogenesis and functional maturation of organoids and can greatly improve the efficiency of organoid preparation.

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.

The invention provides a bovine embryo refrigerated preservation solution. The bovine embryo refrigerated preservation solution is prepared from hydroxyethyl starch, dimethyl sulfoxide, fetal calf serum, cane sugar, cepharanthine and deionized water. The ratio of the components is further optimized, and the components comprise 5% of hydroxyethyl starch, 5% of dimethyl sulfoxide, 55% of fetal calf serum, 37% of deionized water, 2% of cane sugar and 1% of cepharanthine. Under the optimal raw material ratio condition that the concentration of cepharanthine is 600 [mu] mol / L and the concentration of cane sugar is 2 mol / L, cepharanthine is added into the bovine embryo preserving fluid, through a triple synergistic mechanism of oxidation resistance, apoptosis resistance and immunoregulation, the defect of biochemical damage protection in the prior art is overcome, and full-period protection is provided for embryos.

The invention relates to the technical field of biological medicine, and particularly discloses a recombinant bovine interferon coated sustained release microsphere and a preparation method and application thereof, an ultrasonic re-emulsification method is used for replacing bare-handed re-emulsification, 2.5% of trehalose is added into an internal water phase W1, and the protein concentration of the internal water phase W1 is reduced by 30 mg / ml to 25 mg / ml. According to the method provided by the invention, the encapsulation efficiency reaches 90% or above, the recovery rate reaches 95%, the drug loading rate is close to 5%, the balling rate exceeds 90%, and the in-vitro effective slow release time reaches about 10 days; through clinical supplement of the coated slow-release rbIFNT, the pregnancy rate of the receptor cattle in the initial test and the re-test is obviously improved by about 10%, and the pregnancy rate of the repeat breeding infertility young cattle in the initial test is obviously improved by about 15%.

The utility model provides a yak embryo sterile preservation room, which relates to the technical field of sterile preservation and comprises a sterile room body consisting of four groups of coamings, an upper sealing plate and a lower sealing plate. A tooth type conveying belt is rotationally arranged in the middle of each movable display stand, a control motor is fixedly arranged outside each movable display stand, and the control motors are in transmission connection with rotating shafts of the tooth type conveying belts; a pick-and-place channel is fixedly arranged in the middle of the front side of the sterile room body; a bottom guide row is fixedly arranged in the middle of the bottom of the sterile room body, and a vertical sliding frame matched with the guide rail is slidably arranged at the top; an arc-shaped clamping base is arranged, a three-axis moving storage function is provided, through three-axis moving control, storage vessels can be automatically grabbed and fixed and placed above a conveying belt, then a control motor is started to move the tooth type conveying belt by one station, the storage vessels can continue to be placed, and large-scale unmanned storage is achieved; and the storage vessel can be taken out through reverse operation, control is convenient, and the problem that an existing storage chamber is inconvenient to store automatically is solved.

The utility model belongs to the technical field of cattle embryo refrigeration preservation, and particularly relates to a cattle embryo refrigeration preservation device which comprises a liquid nitrogen tank, the upper end of the liquid nitrogen tank is in contact with a tank cover, the outer side of the end of the tank cover is fixedly sleeved with a sealing ring, and the sealing ring is connected with the liquid nitrogen tank in a sliding mode. Two symmetrically-distributed connecting rods are fixedly connected to the end of the tank cover, a fixing frame is fixedly connected to the lower ends of the two connecting rods together, an embryo tank is fixedly connected to the bottom of the inner side of the fixing frame, and a supporting frame is fixedly connected to the outer side of the liquid nitrogen tank. According to the scheme, by arranging a motor, a gear, a toothed plate and other structures, a tank cover can be driven to move so as to drive a fixing frame to move, then an embryo tank is driven to move, automatic taking and using are conducted on the embryo tank, and under the action of clamping holes, clamping rods, first springs and other structures, movement limiting treatment can be conducted on the tank cover; and the embryo tank can be accurately positioned and used.

The invention discloses a method for identifying a pluripotent state of a cattle epidermal layer, and achieves the technical effect of identifying the pluripotent state of a cattle embryo, especially the pluripotent state of the cattle epidermal layer. The invention specifically discloses gene markers for different pluripotent stages and different developmental stages, and the pluripotent and developmental stages of to-be-detected bovine embryonic cells or cell clusters can be rapidly identified by using the gene markers for different pluripotent stages and different developmental stages of bovine embryos.

The invention relates to the technical field of biological information processing, in particular to a cattle embryo pre-implantation genetic evaluation method and system based on whole genome sequencing, and the method comprises the following steps: obtaining whole genome sequencing data of a male parent and a female parent, embryo trophoblast live detection sequencing data and culture solution free desoxyribonucleic acid sequencing data; performing variation detection and haplotype phasing on male and female parent data to form haplotype data, and calculating a haplotype transmission posterior at an anchor point based on trophoblast data; performing genome segmentation based on reading depth observation and allele frequency observation of a trophoblast and a culture solution, and performing Bayesian inference by combining haplotype transfer posteriori as priori to obtain posteriori probabilities of aneuploid, copy number variation, chimera, pathogenic homozygosis and pollution events; and outputting a passing / rechecking / elimination conclusion, adding sequencing and updating the posteriori during rechecking until ending, and outputting a final conclusion and a genome breeding value, so that the evaluation reliability and the decision interpretability are improved.

This invention discloses a kit for amplifying the whole genome of bovine embryonic cells and its applications. The invention provides a primer set for amplifying the whole genome of bovine embryonic cells, consisting of 20 primers, each 6 nt in length. This invention can utilize 4-8 bovine embryonic cells or genomic DNA down to the pg level to amplify and generate whole-genome DNA amplification products with high coverage and low mismatch rate. These products can be directly adapted to downstream applications such as next-generation sequencing and targeted capture sequencing, while reducing detection costs. Furthermore, the three-step operation process of this invention (cell lysis-lysis termination-isothermal amplification) can complete efficient amplification within 3.5 hours.

This invention relates to the field of bovine embryonic stem cell culture technology, and particularly to a long-term culture system for morphological bovine embryonic stem cells completely without a feeder layer and its applications. After trying various culture methods, this invention establishes a feeder-free bovine morphological embryonic stem cell line (FF-bESCs). Bovine morphological embryonic stem cells cultured under the feeder-free conditions provided by this invention can maintain stable proliferation capacity and pluripotency over a long period. The bovine morphological embryonic stem cells provided by this invention can undergo gene editing and can be effectively induced to differentiate into primordial germ cell-like cells (PGCLCs). This invention lays an important foundation for the standardized and large-scale culture of bovine embryonic stem cells under feeder-free conditions, and for their application in stem cell breeding and other fields.

This disclosure relates to methods, products, and compositions useful for inducing bovine induced blastoids. Methods enabling naive stem cell aggregation and induced blastoid formation are described. Aggregation and induced blastoid media suitable for promoting naive stem cell aggregation and preparing induced blastoids are also provided. Bovine induced blastoids may be used in selection and breeding programs, including the replication of pre-transplant embryos with desirable characteristics, as well as in vitro breeding, including cross-generation breeding cycles from parental germ cells and / or embryos to offspring embryos.

The invention belongs to the technical field of embryo transplantation, and particularly relates to a cervix dilation device for bovine embryo transplantation, which comprises a handheld part, a dilation part, a wiping mechanism, a liquid supply mechanism and a storage mechanism, the handheld part comprises a mounting shell and a storage box fixedly mounted at the bottom of the mounting shell; the expansion mechanism comprises a supporting pipe, a control wheel and an extrusion wheel, a through hole allowing the supporting pipe to slide is formed in the outer wall of the mounting shell, the extrusion wheel is used for extruding the supporting pipe and elastically arranged in the mounting shell, the control wheel is rotationally mounted in the mounting shell, and the radial outer wall of the control wheel is attached to the outer wall of the supporting pipe; the wiping mechanism comprises a mounting ring and a wiping layer, the mounting ring is of a hollow structure and is rotatably mounted on the inner wall of the mounting shell, and the wiping layer is fixedly mounted on the inner wall of the mounting ring; the liquid supply mechanism is used for providing disinfectant for the inner cavity of the mounting ring; the storage mechanism comprises a storage box and an elastic strip; through cooperation of the structure, the maternal stress reaction is effectively reduced, and the embryo transplantation success rate and the operation safety are improved.

The application belongs to the technical field of cell biology, and particularly relates to a feeder-free and serum-free bovine embryonic stem cell culture medium, a culture system and a culture method. The bovine embryonic stem cell culture medium comprises a conditioned medium and an additive. The additive is IWR-1 and activin A. The concentrations of IWR-1 and activin A in the conditioned medium are 2.5-5 muM and 20 ng / mL-25 ng / mL respectively. The culture system comprises the bovine embryonic stem cell culture medium, 0.1%-0.2% gelatin in mass concentration and a subculture medium. The 0.1%-0.2% gelatin in mass concentration is used to provide an attachment matrix for the bovine embryonic stem cell. The subculture medium is prepared by adding Y27632 to the bovine embryonic stem cell culture medium. The application reduces the cost of extracellular matrix, and the feeder-free bovine embryonic stem cell has high single cell subculture efficiency and is convenient for gene editing operation.

The application discloses a culture solution for improving the blastocyst rate and quality of bovine embryos, application and an in-vitro culture method, and relates to the technical field of animal embryo engineering. The application provides a culture solution for in-vitro culture of bovine embryos, which comprises a first culture solution and a second culture solution with different osmotic pressure concentrations. By controlling the osmotic pressure concentrations of the first culture solution and the second culture solution in a suitable range, and by using the first culture solution in the early stage of embryo development and the second culture solution in the late stage of embryo development, the in-vitro culture system can better simulate the environment in the oviduct in the early stage of embryo development and better simulate the environment in the uterus in the late stage of embryo development, so that the osmotic pressure stress of the embryo in the development process is reduced, and the blastocyst rate and the blastocyst quality of the bovine embryo are improved. Compared with a traditional culture system with constant osmotic pressure, the application significantly improves the blastocyst rate by sequential regulation of the osmotic pressure, the total cell number of the obtained blastocysts is more, and the apoptosis index is lower.

This invention belongs to the field of biotechnology, specifically relating to an epigenetic regulation inhibitor, a culture medium, and their applications. The epigenetic regulation inhibitor is composed of 1 nM-10 nM of 3-Deazaneplanocin A, 1 nM-10 nM of Trichostatin A, and 10 nM-100 nM of 5-Azacytidine. This invention alters the epigenetic modification state of bovine embryonic stem cells by synergistically regulating histone modification and DNA methylation levels, thereby promoting the activation of pluripotency gene networks and improving the pluripotency and stability of bovine embryonic stem cells. The epigenetic regulation inhibitor of this invention is suitable for long-term culture, maintenance of pluripotency, and preservation of differentiation potential of bovine embryonic stem cells, and can be used in fields such as research on the regulatory mechanisms of bovine embryonic development, screening of important functional genes, and preparation of gene-edited animals.

Differentially expressed miRNA or small mRNA in the culture media of embryos were found to correlate to and affect embryo developmental fate. Accordingly the present invention provides a method for selecting a bovine embryo for implantation into a female bovine animal for further development based on the levels of specific miRNA or small mRNA. Also provided are methods of improving bovine embryo development fate by reducing in the culture medium the level of selected miRNAs.

The invention discloses a kit for predicting the development state of a bovine embryo and application of the kit to diagnosis before embryo implantation. The kit comprises a product for detecting the expression quantity and / or copy number of a CNVR combination 1 in a bovine embryo genome and / or a product for detecting the expression quantity and / or copy number of a CNVR combination 2 in the bovine embryo genome; the CNVR combination 1 comprises eight CNVRs, and the CNVR combination 2 comprises four CNVRs. Experiments prove that a large number of repetitions of CNV fragments of at least one of seven CNVRs in the CNVR combination 1 are beneficial to healthy development of embryos and are potential sites for keeping normal development of the embryos; a great deal of deletion of CNV fragments of at least one of three CNVRs in the CNVR combination 2 is not beneficial to healthy development of embryos and is a potential site causing arrest of embryonic development. The method can be used for scientifically judging the transplantation strategy and decision of the bovine embryo, and has important application value.

The application discloses a kind of embryo active storage devices for bovine embryo transplantation, the liquid nitrogen tank includes shell, inner container and neck pipe, the inner container is located inside shell, the gap between shell and inside is interlayer, the neck pipe is installed at the top of inner container, the top of neck pipe penetrates and extends to the top of shell and is provided with a cap; The placing table is provided with several and circularly arrayed in the inside of inner container, and the adjusting assembly for driving the placing table to move is installed between the side of placing table and the inner wall of inner container. The adjusting assembly moves the placing table to the position directly below the neck pipe, and the bucket is clamped and placed by clamping assembly. The bucket on the placing table can also be taken out by clamping assembly, instead of using frozen semen cloth bag storage and fixed by rope, so that the situation that multiple ropes are wound together and are inconvenient to take out does not occur, effectively avoiding the hand frostbite of staff and the situation of test tube pollution.

The utility model discloses an efficient bovine embryo production incubator which comprises an incubator body, a containing groove is formed in the incubator body, a sealing groove is formed in the incubator body, a fixing rod is fixedly connected to the right inner wall of the sealing groove, a first groove is formed in the fixing rod, and a second groove is formed in the right inner wall of the sealing groove. A first groove is formed in the upper surface of the box body, a placement plate is slidably connected to the inner surface of the first groove, a second groove is formed in the placement plate, an embryo culture dish is arranged on the inner surface of the second groove, and a supporting plate is fixedly connected to the upper surface of the box body. According to the embryo culture dish, mutual interference among different embryos can be effectively avoided, the possibility of cross contamination is reduced, when the embryos need to be taken out, the embryos in other culture dishes cannot be affected, and the embryo culture dish can be limited through cooperation of a pull rod, a first spring, an arc-shaped limiting plate, a second spring and a damper.

The utility model discloses a transfer cabinet for bovine embryo transplantation, which relates to the technical field of transfer equipment and comprises a transfer cabinet body, and a test tube limiting mechanism and a cooling mechanism are arranged in the transfer cabinet body. The test tube limiting mechanism comprises three test tube limiting seats which are distributed in the transfer cabinet body in parallel, and the three test tube limiting seats are rotationally arranged on the inner bottom wall of the transfer cabinet body. According to the transfer cabinet disclosed by the utility model, the test tube limiting mechanism is arranged in the transfer cabinet, so that embryo test tubes can be limited by utilizing the cylindrical slots in the surfaces of the test tube limiting seats, and the test tubes can be ventilated by utilizing the first strip-shaped vent holes and the second strip-shaped vent holes in the surfaces of the test tube limiting seats and the cylindrical slots; cold air generated by the refrigeration box is conveyed into a cylindrical slot by utilizing a flow guide cavity of a test tube limiting seat, the test tubes are cooled by utilizing the cold air, a low-temperature storage effect is achieved, a uniform low-temperature effect can be kept on the surfaces of the test tubes, and transfer and transportation of embryos are facilitated.

The application discloses application of sodium crotonate in improving bovine embryo development potential. In the process of culturing bovine SCNT embryos in vitro, adding sodium crotonate (Nacr) with a proper concentration can promote the development of bovine SCNT cells, significantly improve the development rate and blastocyst rate of the SCNT embryos, improve the embryo quality, and further improve the somatic cell nuclear transfer efficiency of pre-implantation embryos. On this basis, the inventors construct a bovine SCNT early embryo in-vitro culture system, optimize the formula of the in-vitro embryo culture solution and the concentration of Nacr, and further establish a method for improving the development potential of early bovine embryos in vitro. The application can improve the development quality of bovine SCNT embryos, provides a research basis for the rapid propagation of high-yield dairy cattle, high-quality beef cattle and even disease-resistant fine breeds, provides a reference for the batch production of high-quality bovine embryos in vitro, and has important practical significance in the field of SCNT embryo in-vitro culture.

The invention discloses a beef cattle embryo ultralow temperature biological preservation method based on gradient cooling control, and belongs to the field of beef cattle artificial breeding. The mixed solution comprises an equilibrium solution and a cryoprotectant, the equilibrium solution comprises a base solution and fetal calf serum, and the base solution comprises trehalose and polyvinylpyrrolidone; the cryoprotectant comprises the following components: dimethyl sulfoxide, glycerol and antifreeze protein, and the cryoprotectant accounts for 0.5-2% of the total volume of the mixed solution. And balancing the bovine embryos in the mixed solution for 10-15 minutes, and then carrying out ultralow-temperature preservation by adopting a gradient cooling mode. According to the method, the permeation protective agent and the non-permeation protective agent are combined for use, the formula components and the dosage of the permeation protective agent are optimized, meanwhile, a gradient cooling mode is combined, in the stage of starting to form ice crystals, a low-speed rotation mode is combined with gradient cooling, so that the formation of the ice crystals is remarkably reduced, and the embryo preservation effect is improved.

The application relates to the technical field of embryo culture, and discloses a biological-sensing-based optimization method for in-vitro culture of bovine embryos. The method acquires basic culture parameters for in-vitro culture of bovine embryos, sets a monitoring period, and collects biological sensing data in each period; the sensing data are processed to obtain a metabolic fluctuation amplitude, and the basic culture parameters are combined to calculate a culture environment optimization coefficient of each period; the optimization coefficient is compared with a preset safety threshold value, when the optimization coefficient is higher than the threshold value, the culture environment parameters are dynamically adjusted according to the metabolic fluctuation amplitude; the adjustment process is recorded, and when the adjustment times reach a preset threshold value, a warning is triggered; after the culture period ends, the environmental changes are continuously monitored and maintenance operation is performed; key parameters are monitored in real time, and when the key parameters exceed a safety range, emergency intervention is performed; finally, the basic culture parameters are updated according to the adjustment process and the emergency intervention result. The method realizes accurate adaptation of the culture environment, and guarantees the stability of the in-vitro culture process of bovine embryos.