85 results about "In vitro maturation" patented technology

The invention provides a porcine oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the field of embryo engineering, and the additive contains taurine. The additive also contains sodium pyruvate, cysteine, glucose, NaHCO3, fetal calf serum, estradiol, pregnant horse serum gonadotropin and human chorionic gonadotropin. When the porcine oocyte in-vitro maturation culture solution added with the additive is used for culturing porcine oocytes in vitro, the ROS content in the porcine oocytes can be remarkably reduced, the oxidation resistance of the porcine oocytes can be improved, and the mitochondrial distribution of the oocytes can be improved, so that the development of the oocytes can be improved; the maturation of porcine oocytes and the development of early embryos are promoted. When the technology is applied to the pig assisted reproduction technology, the success rate and quality of embryo engineering can be improved; the pig oocyte in-vitro maturation culture solution added with the additive can improve the premature embryo division rate in parthenogenetic activation of the pig oocytes.

The invention relates to an application of a regulator in improving the maturation quality of frozen pig oocytes, the regulator is N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyryl, and the regulator improves the maturation quality of the frozen pig oocytes by improving the mitochondrial function of the frozen pig oocytes after in-vitro maturation. The invention provides a new application of N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyramide, namely the N-(2-(2-(4-chlorphenyl) thiazole-4-yl) ethyl) butyramide is used for developing a novel oocyte vitrification freezing solution and an in-vitro maturation culture solution, and the obtained high-quality oocytes can be beneficial to embryo biotechnology researches such as in-vitro fertilization and somatic cell nuclear transplantation.

The invention discloses a method for optimizing the maturation effect of cattle and sheep oocytes through CNP pretreatment, and relates to the technical field of biological breeding, the method comprises the following steps: S1, selecting and treating donor cells; s2, collection of oocytes; s3, in-vitro maturation culture of the oocytes: after the pre-maturation treatment, transferring the oocytes into a maturation culture solution for continuous culture under the in-vitro culture condition of air containing 5% CO2, keeping the culture temperature of the oocytes within 1-20 hours at 38.5-39 DEG C, and keeping the culture temperature of the oocytes within 20-28 hours at 39-39.5 DEG C; s4, culturing and screening reconstructed embryos; s5, preparing and transplanting a receptor animal; and S6, performing subsequent monitoring and management. According to the method, the culture temperature is kept at 38.5-39 DEG C within 1-20 hours, the culture temperature is kept at 39-39.5 DEG C within 20-28 hours, a series of enzymes and signal channels in the oocytes can be effectively activated by accurately controlling the culture temperature, and the survival rate and the development quality of the oocytes are improved.

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.

The present application relates to the field of animal reproduction, and discloses a method for improving the efficiency of in vivo oocyte collection of dairy goats by using aminophylline, which comprises the following steps: step one, superovulation treatment of dairy goats, (1) placing a vaginal suppository; (2) orally administering aminophylline; (3) injecting follicle stimulating hormone (FSH); step two, an in vivo oocyte collection operation method (LOPU) of dairy goats; step three, in vitro culture of oocytes, in vitro maturation (IVM) of oocytes; in vitro fertilization (IVF); and in vitro culture (IVC) of oocytes.The present application has the advantage that, compared with the prior art, the number of oocytes collected from dairy goats is significantly increased by aminophylline treatment, and the problem of low oocyte collection rate is solved.

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.

This invention discloses an in vitro maturation culture medium for ferret oocytes and its preparation method, belonging to the field of animal embryo engineering technology. It comprises the following components: TCM199 basal culture medium, 50 mL fetal bovine serum, 25 IU follicle-stimulating hormone, 25 IU luteinizing hormone, 0.5 mg 17β-estradiol, 0.005 mg epidermal growth factor, 12.1 mg sodium pyruvate, 0.2 mM cysteine, 0.1 mM cysteine, 0.030 g penicillin, 0.025 g streptomycin sulfate, as well as an antioxidant complex, a combination of growth factors, and extracellular matrix components. This invention provides a comprehensive and suitable nutritional environment for ferret oocytes by precisely formulating a composite culture medium containing multiple key components. Specifically, the hormone combination at specific concentrations accurately simulates the in vivo physiological environment, effectively regulating the oocyte maturation process; multiple growth factors work synergistically to promote oocyte growth, development, and differentiation; and the addition of the antioxidant complex effectively eliminates free radicals generated during in vitro culture, reducing oxidative stress damage to oocytes.

The present disclosure provides compositions, kits, and methods for promoting in vitro maturation of cells. The compositions, kits, and methods utilize at least one inhibitor of an epigenetic regulator and at least one agonist of a calcium channel. Suitable applications of the present disclosure include use in maturing and differentiation stem cells into viable mature neuronal cells for use in disease modeling, prevention and treatment. The present disclosure also provides methods of screening compounds that are suitable for promoting in vitro maturation of cells.

The present application relates to the technical field of biological medicine, and more particularly to a culture solution, a kit and an application for in-vitro maturation of aged oocytes. Through in-vitro experiments, the present application adds exogenous stromal cell-derived factor 1, establishes a culture system for in-vitro maturation of aged oocytes, and reveals the molecular mechanism of stromal cell-derived factor 1 playing a role through autophagy, thereby providing a new solution for improving the fertility of aged women and improving the success rate of assisted reproductive technology, and having important clinical application value.

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 present invention relates to a culture medium composition for in vitro maturation of oocytes, comprising piperine as an active ingredient, and an in vitro embryo production method using same. The culture medium composition for in vitro maturation of oocytes, comprising piperine as an active ingredient, of the present invention, can qualitatively improve oocytes by inhibiting reactive oxygen species, and in vitro embryo production efficiency can be increased using in vitro matured oocytes obtained from the medium of the present invention.

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 provides application of an RTL6 gene in improvement of an in-vitro embryo development rate, and relates to the technical field of mammal embryo engineering. According to the present invention, the RTL6 gene is constructed into the PcDNA3.1 (+)-RTL6 recombinant vector, and the pcDNA3.1 (+)-RTL6 recombinant vector is injected into the in vitro mature oocyte cytoplasm 4-5 h after the pig parthenogenetic activation, such that the embryonic development retardation phenomenon is effectively relieved, and the embryonic development is promoted;

Provided herein are methods for large-scale in vitro maturation of cardiomyocytes derived from human pluripotent stem cells, compositions prepared by these methods, and use of these compositions in cardiac regeneration.

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

Featured are methods, compositions, and apparatuses for the in vitro maturation of oocytes. In particular, the disclosure features methods of inducing oocyte maturation in vitro, by co-culturing a female subject's oocytes with an ex vivo composition containing a plurality of ovarian support cells (e.g., granulosa cells). Additional methods for administering follicular triggering agents and retrieving oocytes from the female subject are provided. Such methods, compositions, and apparatuses are particularly useful for assisted reproduction technology (ART) procedures.

The invention belongs to the technical field of cell biology, and provides an oocyte in-vitro maturation culture solution additive and application thereof. Experimental results show that coumarin 106 can improve the retardation of the meiosis process of oocytes, reduce the form proportion of abnormal spindles of oocytes and the proportion of wrongly arranged chromosomes, recover the level of acetylated tubulin and improve the stability of microtubules; the actin expression level in the spindle body of the oocyte is improved; the conditions of wrong distribution and expression decline of mitochondria in oocytes are improved; the mitochondrial membrane potential level is improved; fluorescence signals of oocyte cortical particle and specific cortical particle protease are improved; the fertilization rate is improved; the active oxygen level and the DNA damage level in the oocytes are reduced, and the occurrence of early apoptosis in the oocytes is reduced. A culture solution prepared from the coumarin 106 can be used for promoting in-vitro maturation of oocytes.

The invention belongs to the technical field of cell engineering and gene engineering, and discloses a method for realizing precise editing of a single allele of an oocyte of herbivorous livestock and application, and the method for realizing precise editing of the single allele of the oocyte comprises the following steps: preparing sgRNA for target oocyte gene editing; carrying out cytoplasm microscopic co-injection on the oocytes; and carrying out in-vitro fertilization and detection on the gene editing oocytes. According to the method, the herbivorous livestock oocyte monoalleles can be accurately deleted. According to the method for specifically knocking out the oocyte gene, sgRNA and Cas9mRNA capable of specifically modifying the oocyte gene in a targeted manner are co-transfected to an oocyte, so that the oocyte gene is knocked out; the co-transfection mode specifically can be microscopic co-injection; the oocytes can be specifically sheep oocytes, and the co-injection time of the oocytes can be 20 hours of in-vitro maturation.

The invention discloses a method for promoting in-vitro maturation of sheep oocytes. The composition comprises an induction culture solution A and an induction culture solution B. The induction culture solution A and the induction culture solution B comprise a basic culture solution, and the induction culture solution A further comprises AMH; the induction culture solution B also contains FSH; the basic culture solution comprises an M199 basic culture medium, oestrus sheep serum, sodium pyruvate, luteinizing hormone, estradiol and a penicillin and streptomycin mixed solution. According to the composition and the method, through the synergistic effect of FSH treatment and AMH treatment, the maturation rate, the activity and the development potential of sheep oocytes are remarkably improved, so that the success rate of in-vitro fertilization and embryo transplantation is improved, an important scientific basis is provided for sheep reproduction and assisted reproduction technologies, related technologies are further optimized, and the application prospect is wide. The reproduction success rate is improved.

The application belongs to the technical field of cell biology, and provides an oocyte in vitro maturation culture medium additive and application thereof. The experimental results of the application show that coumarin 106 can improve the meiosis process blockage of oocytes, reduce the morphological proportion of abnormal spindle and the proportion of misaligned chromosomes, restore the level of acetylated tubulin, improve microtubule stability, improve the expression level of actin in the spindle of oocytes, improve the misdistribution and expression decrease of mitochondria in oocytes, improve the level of mitochondrial membrane potential, improve the fluorescence signal of cortical granules and specific cortical granule proteases in oocytes, improve the fertilization rate, reduce the level of reactive oxygen species and DNA damage in oocytes, and reduce the occurrence of early apoptosis in oocytes. The culture medium prepared by using coumarin 106 can be used for promoting oocyte in vitro maturation.

Featured are methods, compositions, and apparatuses for the in vitro maturation of oocytes. In particular, the disclosure features methods of inducing oocyte maturation in vitro, by co-culturing a female subject's oocytes with an ex vivo composition containing a plurality of ovarian support cells (e.g., granulosa cells). Additional methods for administering follicular triggering agents and retrieving oocytes from the female subject are provided. Such methods, compositions, and apparatuses are particularly useful for assisted reproduction technology (ART) procedures.

The invention belongs to the technical field of biology, and particularly relates to a method for improving in-vitro mature oocyte telomere length and maturation quality and application of the in-vitro mature oocyte telomere length and the maturation quality. And daphnetin. According to the preparation disclosed by the invention, at least one of the Sema3 type protein and the daphnetin is added, so that the telomere length of an oocyte and an in-vitro embryo is effectively maintained, repaired or prolonged, the maturation quality of the in-vitro oocyte and the development potential and development efficiency of the in-vitro embryo are further remarkably improved, and further, the development potential and development efficiency of the in-vitro embryo are further improved. The application provides a more effective new strategy for in-vitro embryo production of livestock, animal genetic improvement, assisted reproduction technology and the like, and the application prospect is wide.

The invention discloses a method for improving in-vitro maturation quality and cryopreservation effect of porcine oocytes, and relates to the technical field of biology. By adding the NMN, the comprehensive efficiency of in-vitro maturation and cryopreservation of the porcine oocytes is remarkably improved. Research finds that by adding NMN into an IVM culture medium, maturation of cell nucleuses is promoted, discharge of first polar bodies of oocytes is improved, oxidative stress and lipid peroxidation of the oocytes are reduced, mitochondrial activity is improved, and cytoplasm maturation of the oocytes is facilitated. Meanwhile, after the NMN is used for pretreatment, the survival rate of the vitrified and frozen oocytes is remarkably improved, and the active oxygen level and the DNA breakage level are reduced. Therefore, the addition of the NMN in the IVM culture solution may become a new method for improving and optimizing the IVM technology, and an innovative solution is provided for pig germplasm resource preservation and excellent breeding stock breeding.

This invention relates to the field of reproductive engineering technology, specifically to the application of rutabagastone in improving the cryopreservation and in vitro maturation of oocytes. This invention is the first to clearly demonstrate that rutabagastone can improve the quality of cryopreservation and in vitro maturation of frozen oocytes. By adding rutabagastone to the cryopreservation solution, thawing solution, and in vitro maturation solution, this invention reduces cryopreservation damage to oocytes, resulting in increased survival rate after thawing, improved mitochondrial function, reduced damage to the oocyte spindle and zona pellucida, reduced oxidative stress levels, and ultimately improved subsequent developmental capacity of frozen oocytes.

The present invention discloses a composition, method and application thereof for improving the quality of in vitro maturation of eggs and the pregnancy rate of embryos after in vitro fertilization. The present invention aims at the problems of low in vitro maturation rate and poor subsequent embryo development potential caused by metabolic disorders during in vitro maturation of oocytes, and provides a composition including two or more regulators in vitamin C, L-carnitine and thrombopoietin to regulate the metabolic mode of oocytes, thereby greatly improving the in vitro maturation rate of oocytes and the subsequent embryo development potential. The present invention not only improves the efficiency of in vitro embryo production of livestock, but also provides a reference for improving the quality of in vitro maturation of human assisted reproductive eggs. The method is simple, safe and efficient, and has great application value and broad application prospects.

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.

Cell culture media comprising luteinizing hormone, follicle-stimulating hormone and at least one histone acetyltransferase inhibitor or reverse transcriptase inhibitor are provided. Methods of in vitro maturing a human oocyte, determining suitability for in vitro maturation, as well as kits comprising media, luteinizing hormone, follicle-stimulating hormone and at least one histone acetyltransferase inhibitor or reverse transcriptase inhibitor are also provided.

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 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.

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