106results about How to "Increase development rate" patented technology

The invention discloses prophase culture solution (foregoing 60h) for porcine early embryonic (applicable to embryos before porcine blastocysts are hatched) development and anaphase culture solution (from 60h after the culture to a blastocyst hatching stage) for porcine early embryonic development. Calcium lactate, sodium pyruvate, beta-mercaptoethanol, dbcAMP.Na, nonessential amino acid and oestrus 7dth porcine serum are added on the basis of NCSU23 culture solution without CaC12. In addition, the difference between the prophase culture solution for porcine early embryonic development and the anaphase culture solution for porcine early embryonic development is that: the prophase culture solution for porcine early embryonic development does not contain glucose, and the anaphase culture solution for porcine early embryonic development contains the glucose; and particularly, when porcine early embryos are cultured, the prophase culture solution for porcine early embryonic development is used for culturing first, and the anaphase culture solution for porcine early embryonic development is used after 60h. Therefore, the developmental rate of the blastocysts and the quality of the blastocysts can be greatly improved.

The invention relates to a mammal embryo in-vitro culture device and a culture method thereof. The culture device is formed by bonding a flow channel layer and a main body layer, wherein flow channels are arranged in the flow channel layer; an electromagnetic coil is arranged below the middle part of the flow channels, and connected with a square-wave generator through a wire; the main body layer is provided with a membrane, and culture chambers are arranged around the membrane and communicated with each other through the flow channels; a magnetic sheet is arranged on the membrane and positioned above the electromagnetic coil; and the wire is provided with a circuit switch. The culture method mainly comprises the following steps: setting frequency of the square-wave generator; injecting a culture fluid and mineral oil required by embryo development into the flow channels from the culture chambers, and putting the in-vitro fertilized embryo into the culture chambers; turning on the circuit switch; and putting the whole culture device into a culture oven containing 5% of carbon dioxide to carry out culture. The invention simulates the mammal in-vivo micro environment of the embryo, stimulates the growth and development of the embryo, enhances the blastula development rate, and increases the cell count of the blastula inner cell mass.

The invention relates to a method for bovine in-vitro fertilized embryo culture and a used culture solution. Particularly, on one hand, the invention relates to a method for culturing a bovine in-vitro fertilized embryo. The method comprises the following steps of putting the bovine in-vitro fertilized embryo into a bovine in-vitro fertilized embryo culture solution and carrying out embryo in-vitro culture under the conditions that the temperature is 38.5 DEG C, the content of CO2 is 0.5% and the humidity is 100%. The invention further relates to a bovine in-vitro fertilized embryo culture solution. The culture solution comprises NaCl, KCl, NaHCO3, MgCl2, KH2PO3, sodium pyruvate, glucose, calcium galactonate, fetal calf serum, L-glutamine, an essential amino-acid, a nonessential amino acid, glutathione and water. The method and the used culture solution have excellent technical effects in the specification, for example, the method has a higher blastocyst hatching rate when the bovine in-vitro fertilized embryo is cultured.

The invention discloses a method for processing bovine somatic cell cloned embryos constructed on the basis of somatic cell nuclear transplantation, which comprises the following steps of: injecting bovine somatic cells to be transplanted into denucleated oocyte to carry out electrofusion; and an hour later after the electrofusion is completed, selecting the successfully fused bovine somatic cellcloned embryos and processing the bovine somatic cell cloned embryos for 12 hours by 1muM of Oxamflatin. By the method, the developmental rate and the quality of the bovine somatic cell cloned embryos can be obviously improved and the bovine somatic cell cloned embryos can be efficiently produced in vitro.

The invention discloses an in vitro fertilization culture dish, which comprises a dish body (1), wherein a first accommodating area (11) for placing ovum and a second accommodating area (12) for placing sperms are arranged inside the dish body (1) and communicated with each other through at least one channel (13) which is tubular; and the top of the channel (13) is provided with a vent hole (15). Moreover, the invention also discloses an in vitro embryo fertilization method for utilizing the culture dish. The in vitro fertilization culture dish and the method can effectively guarantee that only high-quality sperms with enough vigor can reach the ovum and are possible to be combined with the ovum, so as to improve the developmental rate of fertilized embryos and the success rate of embryo transplantation and finally guarantee that bred offspring has superior quality.

The invention provides a method for improving the moveability of sperm in frozen bull semen and the in vitro fertilization rate. The method is characterized in that when SP-TALP washing liquid is used for washing at a prior disposal stage of the frozen bull semen, or when PHE (phenylalanine) capacitation liquid is used for capacitation disposal at a capacitation disposal stage of the semen before fertilization, or when fertilization drips are prepared at an in vitro fertilization stage, glutathione and caffeine of which the purities are both higher than 98% are added; in the fertilization process of each piece of frozen bull semen, in the SP-TALP washing liquid containing bull semen or in the mixed liquid of IVF-TALP seminal fluid containing the bull semen and the PHE capacitation liquid or in the mixed liquid of the fertilization drips containing oocytes and the bull semen, the final concentration of the glutathione and the caffeine is 5mmol/L. According to the method disclosed by the invention, two substances are jointed to be respectively applied to three stages in the fertilization process of the frozen bull semen, the moveability of the sperm is greatly improved, and the fertilization rate of the sperm is improved.

The invention discloses a freezing liquid for preserving embryo, especially a freezing liquid for preserving sampled embryo. The freezing liquid comprises ethylene glycol and sucrose, and is characterized by comprising 0.4-5g of arabinogalactan, 75-85mL of ethylene glycol, 30-40g of sucrose, and 910-930mL of PBS buffer solution containing 0.0021 g/mL bovine serum albumin (BSA). A preparation method of the freezing liquid comprises the steps of dissolving ethylene glycol, sucrose and arabinogalactan in the BSA-containing PBS buffer solution. The freezing liquid fully utilizes synergic effects of the four components that ethylene glycol, sucrose, arabinogalactan and bovine serum albumin are coordinated and supported one another, and has higher development rate of thawed sampled embryo in comparison with conventional ethylene glycol freezing liquid.

The invention provides application of decidua NK cells and decidua NK cell exosomes from cell subsets in preparing drugs and auxiliary therapeutic agents for treating diseases related to infertility.Experiments prove that the exosomes treat endometrium growth disorders by promoting endometrial thickness increasing, improving endometrial cell activity, reducing endometrial cell damage, promoting VEGF expression, maintaining stem features of endometrium matrix cells and stimulating proliferation, so that the pregnancy success rate of endometrium damage model mice is increased to 50-70% from 20%; and the exosomes treat diseases related to maternal-fetal immunologic tolerance disorders by playing a role of immunologic tolerance, reducing a spontaneous abortion rate and improving a help T lymphocyte level. Besides, the exosomes can effectively increase the developmental rate of zygotes fertilized no matter in vitro or in vivo in a multiple mode, meanwhile, an in-vitro fertilization rate, an implantation rate of implantation and a birth rate are also increased, and positive assisting effects on infertility treatment are achieved.

The invention provides a method for improving the in-vitro developmental capacity of ovine oocytes. The method comprises the following steps: carrying out in vitro maturation and in vitro fertilization on inferior BCB-oocytes obtained by chromosome segregation; culturing by 200nMol/L of culture solution containing histone deacetylase inhibitor trichostatin A (TSA) for 10 hours, and continuously culturing by a culture solution without TSA, calculating the cleavage rate after culturing for 48 hours, and calculating the blastocyst rate after culturing for 7 days; and washing the BCB+ oocytes obtained by chromosome segregation by mature culture solution for 2-3 times, and culturing for 24 hours to directly utilize. The method has an important practical value on in vitro utilization of ovine oocytes.

The invention relates to a modified culture method for promoting cloned mouse embryonic development. The method is a continuous culture method-D culture method. The invention also relates to two culture systems capable of increasing the developmental rate of cloned mouse embryo, a method of using the culture systems for obtaining cloning animal and a method of using the culture systems for obtaining embryonic stem cells. By using the culture method of the invention, the developmental rate of cloned mouse embryo and the establishment efficiency of nuclear transfer of embryonic stem cells are increased, thus paving a new way for using clone technology in the production and facilitating the further development of animal cloning technology.

The invention relates to methods for assisted reproduction technology in mammals. Specifically the invention relates to methods for in vitro mammalian oocyte culture and oocyte maturation, in vitro fertilization, and in vitro embryo development.

The utility model discloses a special clamping tool for a robot for the live replacement of a tension single insulator of an ultrahigh-voltage line. Each of a front clamp (3) and a rear clamp (1) comprises a main body (101) and an upper cover (102), wherein a collision block locking mechanism is arranged between the upper cover (102) and the main body (101); a motor locking mechanism (9) is arranged at the threaded connection part of the upper cover (102) and the main body (101); a motor (901) of the motor locking mechanism (9) is electrically connected with a robot controller; an adjusting lead screw (5) and a pulling plate frame assembly (2) are respectively and rigidly connected to two ends of the front clamp (3) and the rear clamp (1) to form an insulator tightening device; the end ofone side of the front clamp (3) and the end of one side of the rear clamp (1) are fixedly connected with a robot arm (7). The clamping tool has the advantages that the clamping tool is suitable for arobot to realize automatic replacement of the low-value single insulator at high altitude, the structure is simple, and automatic operation is easy to realize.

The invention belongs to the field of assisted reproduction and particularly relates to vitrification refrigerating liquid. According to the vitrification refrigerating liquid provided by the invention, taxol is added to serve as a cryoprotectant and the vitrification refrigerating liquid obtained with reference to the taxol adding mode is treated, so that the development rate of in vitro fertilized blastocyst after the oocyte is thawed can be increased. The taxol serving as an additive in the refrigerating liquid can achieve the effects of reducing freezing injury of the mature oocytes and improving the development capability of the embryo after fertilization. The optimal concentration of the adding mode is obtained by researching the adding concentration of the taxol. The vitrification refrigerating liquid provided by the invention can greatly increase recovery rate after the frozen mature oocytes are thawed and the development rate of the blastula and achieves the protective effecton the cell ultrastructure.

The invention provides a method for improving compatibility and development rate of interracial nuclear transfer embryo nucleoplasm. The method includes the steps: (a) a cell nucleus of a nuclear donor cell is moved into a perivitelline space of an enucleated oocyte of a non-human mammal and integrated by means of electric stimulation, so that a reconstructed ovum is formed, (b) a human oocyte is moved into the reconstructed ovum in the step (a), and (c) the reconstructed ovum in the step (b) is electrically stimulated so as to form a reconstructed ovum composed of the cell nucleus of a human cell, the enucleated oocyte of the non-human mammal and the human oocyte. The embryo development rate can be increased remarkably by the method.

The invention discloses a low-density high-strength petroleum ceramsite proppant taking tuff as a main raw material, and a preparation method thereof. The ceramsite preparation materials is prepared from the following components in percentage by weight: 40%-60% of tuff mineral powder, 30%-45% of bauxite and 5%-20% of magma powder, wherein the total weight percent is 100%, and the volume of modified polyphenyl particles accounts for 30% of the total volume of the materials. The method comprises the following steps: weighing a certain amount of tuff, bauxite and a small amount of magma, blending and grinding, adding a certain amount of polyphenyl particles in a ceramsite pelletizer, pelletizing through a ceramsite making machine to make balls, drying the balls, screening the balls according to grain size, and roasting the qualified balls at the temperature of 1100-1190 DEG C for 20-40 minutes, so as to obtain the fired low-density high-strength proppant. The low-density high-strength petroleum ceramsite proppant utilizes the characteristics of high silicon content of the tuff and the magma, so that the ceramsite firing temperature is reduced, the ceramsite strength is improved at the same time, and the production cost is lowered.

The invention discloses a vector and method for increasing cattle cloning efficiency based on histone methylation level modification. The vector comprises the in-vitro transcription template vector containing a cattle KDM4E gene. A large amount of mRNA of the cattle KDM4E gene can be obtained by subjecting the vector to in-vitro transcription reaction, and the mRNA is injected to a successfully fused and activated bovine somatic cell cloning embryo to allow the embryo to overexpress cattle KDM4E protein. The method has the advantages that histone H3K9me3 and H3K9me2 epigenetic modification level of the somatic cell cloning embryo during zygotic activation can be effectively lowered, so that the development rate and quality of later bovine somatic cell cloning embryo can be increased evidently, the method can be used for efficiently producing the bovine somatic cell cloning embryo in vitro, and the birth rate of cloning cattle after embryo transplanting is increased evidently.

The invention discloses a method for improving cattle cloning efficiency by micro ribonucleic acid. bta-miR-125b mimic is added into electrical fusion solution of somatic cell nucleus transfer and can be effectively led into an embryo, the histone H3K9me3 epigenetic modification level of a somatic cell cloning embryo in a zygotic activation period can be effectively weakened while secondary damage of transfection is avoided, and the developmental rate and the quality of a cattle somatic cell cloning blastaea can be obviously improved.

A method for producing a monkey-derived embryonic stem cell comprising the steps of carrying out fertilization by insemination by in vitro fertilization or intracytoplasmic sperm injection using a monkey ovum and monkey sperms, thereby giving a fertilized ovum, allowing the fertilized ovum to differentiate into a blastocyst by in vitro culture, and establishing an ES cell line using the blastocyst; the monkey ES cell obtained by the method, a method for screening a reagent for specific differentiation into cell or tissue by using the ES cell; and a differentiated cell or differentiated tissue each differentiated from the ES cell. According to the present invention, applications of the embryonic stem cells to embryological studies clinical applications, experimental models, and the like on primates, studies of diseases, are expected.