53results about How to "Increase blastocyst rate" patented technology

The invention discloses an in-vitro maturating method for sheep oocyte, a pretreatment solution and a kit. The related pretreatment solution applied to in-vitro maturating of sheep oocyte is characterized by comprising sheep small follicle fluid and routine culture-solution compositions for in-vitro maturating of sheep oocyte. The disclosed in-vitro maturating method for sheep oocyte is characterized in that a pretreatment step is increased on the basis of a conventional in-vitro maturating method. Also, experiments in the invention verify that oosperm obtained after an oocyte which is cultured by employing the maturating method is subjected to in-vitro fertilization has the cleavage rate and the blastocyst rate both substantially higher than those of an oocyte cultured by employing a conventional maturating method. The provided pretreatment solution and the in-vitro maturating culturing method are capable of effectively promoting in-vitro maturation of sheep oocyte, and have the characteristics of no hazard to occyte, few limits, good effect and the like.

he invention discloses a method for producing somatic cell cloned bovine blastocysts. According to the method, to bovine ear limbal fibroblasts are used as nuclear transfer donorcells, and mature bovine oocytes cultivated in vitro are used as nuclear transfer receptor cells; the oocytes are denucleated by extrusion and removed with the zona pellucida; the oocytes and the donor cells are adhered in an arrangement way of oocytes-donor cells-oocytes and treated with electro-fusion to obtain reconstructed embryos; and the reconstructed embryos are activated and cultured in vitro to obtain the cloned blastocysts. The method can improve the production efficiency of the cloned bovine blastocysts.

The invention relates to application of n-octanol to the prolonging of the in-vitro ageing of an ovum of a mammal. According to the application, the n-octanol is added in a culture solution containing fetal calf serum, and the concentration of the n-octanol is 0.1-4 mM. By using the application, the speed of the in-vitro ageing of an oocyte of the mammal is obviously deferred; meanwhile, the in-vitro development capability of the oocyte is guaranteed, so as to provide a novel approach for deferring or reversing the ageing of the ovum; and a reference is provided for improving an auxiliary reproductive technology. The technology provided by the invention is not reported in public at home and abroad, has extremely high innovativeness and has a great impellent action on the field of the embryo engineering of the mammal.

The invention discloses a method for improving pig nucleus transplantation efficiency, which belongs to the field of breeding or inseminating methods. The method comprises the following steps of: processing a donor cell by a small molecule medicament Calyculin A and inducing donor chromatin condensation, methylation rise and acetylation reduction, wherein a somatic nucleus expresses genetic modified zero resetting; then carrying out nucleus transplantation and constructing reconstructed embryos; and selecting optimal concentration of 10nM and the processing time of 30 minutes by an optimizing condition. The invention improves the ectogenesis blastocyst rate of a pig clone embryo by 9.7 percent, increases 23.78 blastomeres and lays the foundation for improving the production efficiency of clone animals.

The invention relates to the field of animal husbandry and veterinary and particularly discloses a method for delaying the reproduction function decline of female animals, wherein the animals take in melatonin through oral administration. The delaying of the reproduction function decline of female animals is shown by increasing the litter size of older female animals, the SOD level of ovary, the SOD level of uterus, the number of useful eggs, total egg number, cleavage rate or blastocyst rate or reducing the MDA level of the ovary of older female animals, the MDA level of uterus or the number of useless eggs. In the invention, through a large quantity of objective experiments, different optimal intakes are provided aiming at reproduction function decline of specific different female animals so as to realize the best effect of delaying the reproduction function decline of female animals.

The invention discloses a pig oocyte in-vitro maturation culture solution and a preparation method and application thereof, and belongs to the technical field of oocyte in-vitro maturation culture. The problems of low in-vitro maturation rate and development rate of porcine oocytes currently are solved. The pig oocyte in-vitro maturation culture solution is prepared from a basal culture solution TCM-199, penicillin, streptomycin, NaHCO3, 4-hydroxyethylpiperazine ethane sulfonic acid, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, epidermal growth factor, porcine follicular fluid, pregnant mare serum gonadotropin, human chorionic gonadotropin and tannic acid. According to the pig oocyte in vitro maturation culture solution, the oocyte in-vitro maturation quality, the cumulus cell diffusion index, the parthenogenetic embryo blastocyst rate and blastocyst total cell number, and the in-vitro fertilization embryo cleavage rate, blastocyst rate and blastocyst total cell number can beimproved.

The invention relates to optimization liquid for in vitro maturation of advanced-age mouse oocytes. A preparation method comprises the steps of firstly, weighing 0.0232g of melatonin, enabling the melatonin to dissolve in 1ml of anhydrous ethanol, preparing 100mM of concentrated storage liquid, diluting 100mM of the concentrated storage liquid in an IVM culture medium until the concentration is 10-6mol/L, and the final concentration of the anhydrous ethanol is 0.01% smaller than that of the IVM culture medium, and preparing the optimization liquid for in vitro maturation of advanced-age mouseoocytes. In accordance with in vitro maturation culture liquid for commercial mouse oocytes, the culture efficiency cannot meet production and embryo transplanting requirements, and the in vitro maturation culture liquid for commercial mouse oocytes is improved, so that the maturing rate ( 87.53 +/-2.88% ) and the cleavage rate ( 79.04 +/-3.13% ) after fertilization, and the blastula rate( 65.69 +/-2.07% ) of the advanced-age mouse oocytes can be notably increased. Besides, the distribution and the activity of mitochondrial can be improved, and the oxidation stress level and the early apoptosis are reduced to weaken oocyte degradation caused by aging.

The invention provides a method for establishing a parthenogenetic activation derived embryonic stem cell system, which comprises the following steps: pregnant mare serum gonadotropin (PMSG) is used to act on a female animal body for superovulation; after ova are taken, a calcium ion carrier A23187 and 6-dimethylaminopurine (6-DMAP) are applied to perform chemical activation to the ova, and embryos developed by the activation are cultured in vitro to a blastula; and a mechanical method is used to separate cells of an inner cell mass, a thin needle is used to directly pierce the blastula of the embryos, and the inner cell mass is peeled and is subject to passage and amplification culture in a culture dish paved with feeder layer cells to establish a steady animal parthenogenetic activation ES cell system. The method has good effect of activation treatment, ensures that the blastula rate is up to 96.5 percent, avoids an immunologic reaction from damaging the cells of the inner cell mass by separating the cells of the inner cell mass with the piercing mechanical method, has short time consumption, reduces the exposure time of the cells outside an incubator, furthest protects the cells, and solves the immunogenicity problem in the application of embryo source, namely ES cells.

The invention provides a method for improving in vitro fertilizable competence of vitrification freezing oocyte. The method disclosed by the invention comprises the following steps: performing combined treatment on bovine oocyte with a solution containing gadolinium and CLC before vitrification freezing treatment, and performing vitrification freezing; treating with methyl-beta-cyclodextrin afterunfreezing, and performing in vitro fertilization. Results prove that the method is capable of obviously improving the cleavage rate and blastocyst rate of vitrification freezing bovine oocyte and achieving in vitro fertilization efficiency higher than that of fresh bovine oocyte, has a wide application prospect and is worthy of popularization.

The invention relates to the field of animal cloning methods, in particular to a method for improving the development efficiency of a porcine cloned embryo. The method mainly comprises the step of adding a uterine fluid of a sow into a basic culture solution for the porcine cloned embryo. By the method, the blastocyst rate during early development of the cloned embryo and the total number of cellsin a blastocyst are significantly increased; a transcriptome sequencing result of the early embryo shows that culture through addition of the uterine fluid of the sow reduces the number of differentially expressed genes between the cloned embryo and a normal in-vivo fertilized embryo; most importantly, through the addition of the uterine fluid of the sow, also increased the fertility rate of therecipient sow of the cloned embryos and the number of litters per litter, the parturition rate of a receptor sow with the cloned embryo and the number of live births are also increased, which means that by the method, the final efficiency of the in-vivo development of the cloned embryo is effectively improved.

The invention provides a method for improving the quality and efficiency of in vitro maturation of an oocyte, which includes the step of changing an in vitro maturation fluid at least once during in vitro maturation, and further comprising adding an amount of FGF to the in vitro maturation fluid. The oocyte prepared by the method of the invention has remarkably improved nuclear maturation efficiency and cytoplasmic maturation efficiency, the MII oocyte is up to 95%, the proportion of oocytes with normal distribution of cortical granules and mitochondria, the levels of glutathione and ATP are significantly higher than those of the normal maturation group; Adding a certain amount of FGF to IVM can further improve the maturation quality and efficiency of oocytes. After in vitro fertilization,the in vitro fertilization efficiency of the oocytes prepared by the method of the invention is significantly higher than that of the normal maturation group, which is embodied in higher cleavage rate, blastocyst rate and more blastocyst cells, and has great application value.

The invention provides an oocyte in-vitro maturation culture solution additive and application thereof. The additive is dimethyl alpha-ketoglutarate. The oocyte in-vitro maturation culture solution additive disclosed by the invention is added into an oocyte in-vitro maturation culture solution and is used for culturing oocytes in vitro; according to the method, the maturation rate and quality of the oocytes are remarkably improved, the in-vitro fertilization rate and blastocyst rate are remarkably improved, the ROS content of the oocytes after oxidative damage is remarkably reduced, and the GSH content in the oocytes after oxidative damage is remarkably improved. The success rate and quality of embryo engineering can be improved when the method is applied to a livestock assisted reproduction technology.

The invention discloses a method for improving in-vitro development efficiency of porcine cloned embryos. The method comprises the following steps of: performing molecular identification on bone marrow-derived mesenchymal stem cells, and establishing a method capable of identifying the purity of the cells by using a few cells through a 6-channel flow type technology; performing gene modification on the porcine bone marrow mesenchymal stem cells to obtain porcine bone marrow mesenchymal stem cells for expressing multiple reprogramming factors; and separating out positive porcine bone marrow mesenchymal stem cells by using a fluorescent flow type separation method, continuously culturing the positive porcine bone marrow mesenchymal stem cells for 3 to 7 days, performing nuclear transfer on the cultured cells serving as nuclear donor cells, and studying the influence of 4RFs-3days, 6RFs-3days, 4RFs-7days and 6RFs-7days pMSC serving as nuclear donor cells on the improvement of the development efficiency of the porcine cloned embryos. The results show that the 4RFs-7days pMSC can well promote fission of the cloned embryos and formation of blastospheres, the cloned embryos can form a homogeneous state with uniform cell quantity, and a foundation is laid for efficiently cloning adult high-quality pig varieties in large scale.

The invention discloses an application of an expression inhibitor of a pig SIRT1 gene in preparation of a product with an effect of improving development performance of a pig clone reconstructed embryo. The expression inhibitor of the pig SIRT1 gene can effectively improve the development rate of the pig cloned and reconstructed embryo and the embryo quality in a blastula period, so that the development performance of the pig cloned and reconstructed embryo can be effectively improved, so that the cloning efficiency of porcine somatic cell cloning can be improved. The expression inhibitor of the pig SIRT1 gene can be selected from at least one of siRNA capable of inhibiting the expression of the pig SIRT1 gene and a compound capable of specifically inhibiting the expression of the pig SIRT1 gene. The invention also provides three siRNAs capable of inhibiting the expression of the SIRT1 gene of the pig.

The invention relates to the technical field of buffalo embryo development, in particular to a method for improving the embryo development rate of in vitro fertilization of a river buffalo. Accordingto the present invention, the improvement of buffalo oocyte mature culture solution is mainly researched, and the cleavage rate and blastocyst rate of buffalo fertilized eggs can still be increased inlow concentration of follicular solution (1%-4%), so as to improve the in vitro embryo development rate and produciton efficiency of buffaloes. In the formula, in order to activate the activity of oocytes, the applicant does a large number of studies, and finds that the effect of follicular solution on the promotion of embryo development rate can be significantly improved by dissolving cysteine,methionine, alanine, lysine, tryptophan and glycine in TCM199 solution and then chelating the above solution with the follicular soluiton.

The invention discloses a mouse sperm in-vitro culture kit capable of improving a blastocyst rate and a using method thereof. A current blastocyst rate of mouse in-vitro fertilization is lower, and itis not beneficial to development of assisted reproductive technology research depending on a mouse embryo. The mouse sperm in-vitro culture kit capable of improving the blastocyst rate comprises a mLof mHTF, b mL of BSA, c mg of an ERO1L recombinant protein and d mg of a DPEP2 recombinant protein. a:b:c:d=2:20:1:2. The ERO1L recombinant protein is an endoplasmic reticulum oxidordeuctase 1 albuminoid, and a gene ID in a DNA sequence database is 319446. The ERO1L recombinant protein and the DPEP2 recombinant protein are added to the kit, so the kit is capable of remarkably improving sperm activity and the blastocyst rate.

The present invention discloses a method for the nuclear transplantation. The method is characterized in that donor cells and recipent oocytes of non-human mammals of the same mitochondria haplotype are nuclear transferred. The method in the present invention can remarkably improve fusion, cleavage and blastocyst rates of the reconstructed embryo, thus remarkably improving the nuclear transplantation efficiency.