18 results about "Bovine oocyte" 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.

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 a bovine oocyte cryoprotectant and application thereof, and belongs to the technical field of cell freezing. The method comprises the following steps: firstly, immersing bovine oocytes in vitrification equilibrium liquid, transferring the bovine oocytes into vitrification freezing liquid which contains CspB protein and does not contain DMSO, and utilizing the effects of inhibiting ice crystals and reducing oxidative damage of the oocytes of the CspB protein; then, the treated oocytes are subjected to conventional cryopreservation; after unfreezing, the survival rate of the oocytes is remarkably improved, the problem of toxicity of DMSO to the oocytes is avoided, and the oocytes show better cellular morphology and relatively high development potential at the same time.

The invention relates to the technical field of gene engineering, and discloses a method for obtaining smooth hair surface type dairy cow by injecting RNP (Ribonucleic Acid) into cattle fertilized egg cytoplasm, which comprises the following steps: synthesizing sgRNA (Single Guide Ribonucleic Acid); collecting and maturing bovine oocytes; carrying out in-vitro fertilization and early embryo culture; in-vitro embryo microinjection: placing the fertilized ovum in a micromanipulation solution, sucking a mixed solution of WT-Cas9 protein and sgRNA188 by a microinjection needle, carrying out injection, and after the injection is completed, placing the fertilized ovum in BO-IVC liquid drops for continuous culture; embryo transplantation; birth calves are obtained through delivery, and genotype identification is carried out. According to the method for obtaining the smooth hair surface type dairy cow by injecting RNP into the cattle fertilized egg cytoplasm, the mixed solution of sgRNA188 and Cas9 wild type protein is injected into the cattle fertilized egg, the SLICK phenotype Holstein dairy cow is cultivated, and an effective breeding method is provided for breeding of heat-resistant variety cattle.

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.

The present invention relates to a bovine oocyte cryoprotectant and application thereof, belonging to the technical field of cell freezing. The method comprises: firstly, immersing the bovine oocyte in a vitrification equilibration solution, transferring the bovine oocyte to a vitrification freezing solution containing CspB protein but not containing DMSO, utilizing the CspB protein to inhibit ice crystals and reduce oxidative damage to the oocyte; then, conventionally cryopreserving the treated oocyte; after thawing, the survival rate of the oocyte is significantly improved, avoiding the toxicity problem caused by DMSO to the oocyte, and exhibiting better cell morphology while showing higher developmental potential.

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 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.

The application discloses a one-step freezing solution for bovine in-vitro embryos and a one-step freezing method for bovine in-vitro embryos and belongs to the technical field of biotechnology.The method comprises the following steps: bovine oocyte in-vitro maturation; bovine in-vitro fertilization embryo production; after culturing the bovine in-vitro fertilization embryo for 6-8 days, the embryo is moved into freezing solution A, and after standing, the shriveled blastocyst is moved from the freezing solution A into freezing solution B, the embryo sinking to the bottom of the freezing solution B is moved into freezing solution C, is loaded into a macaroni tube, and is subjected to programmed freezing; after the freezing program is completed, the macaroni tube is put into liquid nitrogen. The bovine in-vitro embryos are frozen by using the three kinds of freezing solutions, on the premise that the conventional freezing method has low technical requirements for the operator and can be directly transplanted after thawing, the conventional freezing method is overcome, the survival rate, the hatching rate and the transplantation pregnancy rate of the embryos after thawing are low, the convenience of use and the effective integration of the high transplantation pregnancy rate are realized.

The invention discloses a culture solution for in-vitro culture of bovine oocytes and application of the culture solution, and belongs to the technical field of livestock embryo engineering. In order to solve the problem that in the prior art, a culture solution for in-vitro culture of bovine oocytes lacks an immune system, so that the in-vitro culture of the oocytes is greatly different from normal development in vivo, a proinflammatory factor IL-1alpha and an anti-inflammatory factor IL-10 secreted by macrophages are added into the culture solution for in-vitro culture of the bovine oocytes; further, the immune requirement in the bovine oocyte in-vitro culture process is perfected, the development quality of bovine oocyte in-vitro culture can be effectively improved, and the embryo development efficiency after oocyte fertilization is improved.

The invention relates to the technical field of in-vitro embryo culture, in particular to a bovine oocyte in-vitro maturation culture solution and application thereof. The bovine oocyte in-vitro maturation culture solution is prepared from Dac51, SAM and zinc sulfate, the ratio of Dac51 to SAM to zinc sulfate is (1-10) [mu] M to (40-200) [mu] M to (0.4-2) [mu] g / mL. According to the bovine oocyte in-vitro maturation culture solution capable of effectively promoting embryonic development, the Dac51, the SAM and the zinc sulfate cooperate to improve the nuclear maturation rate, the cleavage rate, the blastocyst rate, the hatching rate and the freezing survival rate of embryos, and the culture solution has important significance for improving the production efficiency and quality of the embryos.

The present invention relates to the field of in vitro maturation culture of oocytes, and particularly to an additive, a culture medium for promoting the quality of in vitro matured bovine oocytes and their applications; the additive for promoting the in vitro maturation of oocytes is NAD+, and the concentration of the additive NAD+ is 50 μmol / L. A culture medium for promoting the in vitro maturation of oocytes, the components of the culture medium are: M199 medium, 10 mmol / L Hepes, 10% FBS, 0.01 μg / mL FSH, 1 μg / mL LH, 0.01 μg / mL E2, 10-100 μmol / L NAD+. Adding NAD+ to the in vitro maturation culture medium of oocytes can effectively increase the number of mature oocytes and improve the quality of in vitro matured oocytes. By adding NAD+ to the in vitro maturation solution of oocytes, the in vitro maturation efficiency of oocytes can be improved, the existing oocyte maturation culture system can be improved, and then it can be applied to technical fields such as in vitro fertilization, parthenogenetic activation, somatic cell cloning, etc., so as to improve its production and application value.

The invention discloses a method for improving the reproductive performance of cattle by using betaine, and belongs to the technical field of animal reproduction biology. The method comprises applying an effective dose of betaine to bovine oocytes and / or granular cells thereof; the betaine is a natural quaternary ammonium type alkaloid with an anti-oxidation function. According to the invention, it is found through metabonomics that betaine is a key metabolite of cattle high-yield breeding traits for the first time, and it is proved that betaine has double beneficial effects in a cattle breeding system: on a granular cell level, the level of active oxygen in cells induced by hydrogen peroxide can be significantly reduced by adding 5 mM betaine, and oxidative stress can be effectively relieved; on the oocyte level, 0.5 mM betaine is added into an in-vitro maturation and / or embryo culture solution, so that the cleavage rate and blastocyst formation rate of the bovine in-vitro embryo can be remarkably increased. The invention provides a brand-new, safe and effective solution for improving the production efficiency and the reproductive capacity of the bovine in-vitro embryo.

The present application relates to the technical field of gene editing, in particular to a composition for promoting the development of bovine gene editing cloned embryos, a culture solution and application thereof. The present application adds the composition (DLL3, Vercirnon and ML604086) in the in vitro maturation culture medium of oocytes, and effectively reduces the apoptosis and autophagy damage of bovine oocytes through the synergistic cooperation of DLL3, Vercirnon and ML604086, thereby improving the cleavage rate and blastocyst rate of embryos, which has important practical value for improving the production efficiency and quality of gene editing cloned embryos.

The invention provides a method for improving the production efficiency of bovine in-vitro embryos, and belongs to the technical field of livestock breeding biology. According to the method, carnosine is added in the bovine oocyte in-vitro maturation stage, melatonin is added in the embryo culture stage after in-vitro fertilization, and the embryonic development effect is remarkably improved through the staged synergistic effect. According to the method, the characteristics of energy metabolism regulation and control of carnosine and oxidation resistance of melatonin are fully utilized, the defects of oxidative stress and insufficient energy supply in in-vitro culture are effectively overcome, the blastocyst development rate and the embryo quality are improved, and the method is high in safety, easy and convenient to operate and good in application prospect.

The invention discloses a culture solution and a culture method for promoting in-vitro maturation of bovine oocytes. The culture solution for promoting in-vitro maturation of the bovine oocytes comprises a bovine oocyte in-vitro maturation solution and BDNF, and in the culture solution, the concentration of the BDNF is 10-100 ng / mL. The culture method for promoting in-vitro maturation of the bovine oocytes comprises the following steps: adding the oocytes into the culture solution for promoting in-vitro maturation of the bovine oocytes according to the claim 3, and culturing. The invention further provides application of the BDNF in promoting in-vitro maturation of bovine oocytes. According to the method, the in-vitro maturation rate of the bovine oocytes can be remarkably increased, correct distribution of cortical particles and mitochondria can be remarkably promoted, the copy number of the mitochondria can be remarkably increased, and the mitochondrial membrane potential level of the oocytes can be remarkably improved, so that the quality of the in-vitro mature bovine oocytes can be improved, and the early embryo development capability can be improved.

The invention discloses a bovine oocyte negative pressure suction device and a control method thereof.The bovine oocyte negative pressure suction device comprises a negative pressure suction device shell, a control module is arranged in one end of the negative pressure suction device shell, a negative pressure generation module and an operation display module are arranged beside the control module in a connected mode, and the negative pressure generation module is connected with an oocyte suction device; the control module, the negative pressure generation module, the operation display module, the oocyte suction device and the heating test tube rack are arranged in the negative pressure suction device shell, the control module, the negative pressure generation module, the operation display module and the oocyte suction device are arranged in the heating test tube rack, and the heating test tube rack is arranged in the negative pressure suction device shell. Therefore, accurate and stable negative pressure suction of the oocytes and safe storage of the oocytes in a constant-temperature test tube accurate temperature environment are achieved, high control accuracy and working efficiency are achieved, the liquid crystal screen accurately displays set and real-time working temperature and negative pressure values, data are visual and clear, user operation is easy and convenient, and the product is practical in function and high in reliability.

The present invention provides a method for improving the efficiency of bovine in vitro embryo preparation, comprising the steps of in vitro collection of bovine oocytes, in vitro maturation and culture of cumulus-oocyte complexes, in vitro fertilization of bovine oocytes, and in vitro development and culture. The culture medium used in the in vitro development and culture contains 0.01-70 μM C 35 H 49 F2N7O4·CF3CO2H (MM102). By selecting the optimal bovine IVF system under in vitro conditions and placing the blastomeres in development medium supplemented with varying concentrations of MM102 after 48 hours of IVF development, the results showed that MM102 promoted the in vitro development of bovine zygotes, resulting in a high blastocyst rate. This method can be used to improve the efficiency of bovine IVF embryo preparation. This invention can provide a theoretical basis for research on livestock IVF technology and can be further applied to livestock breeding and genetic conservation.