42 results about "Fertilised ovum" 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.

The invention discloses a high-survival-rate sheep fertilized ovum culture method for embryo in vitro culture. The method comprises the following steps of oocyte culture, seminal fluid treatment, in vitro fertilization and fertilized ovum culture, wherein the fertilized ovum culture needs to be performed by putting fertilized ova into a culture box; and the culture box comprises a box body, a plurality of culture vessels put in the box body, an ascending and descending device used for singly ascending and descending the plurality of culture vessels, a control device for controlling the temperature, CO2 and O2 in the box body, and an isolation mechanism for realizing the isolation from the box body in the culture vessel taking or storage process. The high-survival-rate sheep fertilized ovumculture method for embryo in vitro culture has the advantages that the survival rate of the fertilized ovum is improved; and through the arrangement of the culture box, the nutritional environment inthe culture box can be maintained in the process of continuously checking the fertilized ovum.

The present invention proposes formative agent of protein complex, in which a polyphenol is useful component, and the agent is useful as gene complex, cell adhesion inhibitor or immune tolerogen. The polyphenol of forming the agent is selected from catechin group consisting of epigallocatechin-gallate, tannic acids, or proanto-dianisidine, a protein of the protein complex is selected from proteins consisting of animal proteins composed of polypeptide chain of peptide-combined amino acids, vegetative proteins, nucleus proteins, glycogen proteins, lipo-proteins and metal proteins, the gene complex comprises by compositing genes by polyphenol catechins in order to introduce genes to cells of animals or human bodies, a cell composed of the cell adhesion inhibitor is selected from cells consisting of an animal cell including a stem cell, skin cell, mucosa cell, hepatocyte, islet cell, neural cell, cartilage cell, endothelial cell, epidermal cell, osteocyte or muscle cell isolated from human or animal organism, or sperm, ovum or fertilized egg of domestic animals or fishes and a tissue or an organ for transplantation of the immune tolerogen is selected from the tissue or the organ consisting of skin, blood vessel, cornea, kidney, heart, liver, umbilical cord, bowels, nerve, lung, placenta or pancreas.

A mouse animal model C57BL / 6 for the APP gene of human presenile dementia (AD) is disclosed, which can be used to research the AD, develop its medicine and screen its vaccine. Its configuring process includes using PDGF promoter to regulate APP and configure its transgene, injecting it in the male pronucleus of fertilized ovum of mouse, PCR, southern discrimination to obtain transgenic male animal, cesarean section, and DNA recombination-inbred-line passage for configuring its line.

The invention discloses a navodon septentrionalis spawning site and an ovum harvesting method. A spermatovum harvesting device of the navodon septentrionalis spawning site comprises a plastic corrugated sheet which is taken as a base of the device, lead weights or other weights are arranged on four corners of the base of the device, and four pieces of rectangular soft plastic cloths which are mutually staggered are arranged on the base of the device. The device is placed at the bottom of the navodon septentrionalis spawning site, the soft plastic cloths which are sewn and fixed on the device are placed upward, the direction of the soft plastic cloths is vertical toward the direction of water flow, and ova adhered on the plastic corrugated sheet base and the soft plastic cloths are collected after parent fishes spawn. The device can simulate condition of the navodon septentrionalis spawning site in natural sea area, an environment which is more familiar and adaptive to the parent fishes is formed, the parent fishes can be shielded, and stress response in a parent fish rearing process is reduced, thus being more convenient for the mating and spawning of the parent fishes.

The invention relates to an expression vector containing a calpain-1 gene. The expression vector comprises a Tet operon, an alpha-MHC promoter nucleotide sequence, a mitochondrial signal peptide sequence and a gene segment of calpain-1 which are connected in sequence. The invention further provides a construction method of an inducible transgenic mouse cardiomyopathy animal model. The constructionmethod comprises the following steps: importing the expression vector containing the calpain-1 gene into oosperms of a mouse, and transferring the oosperms into the uterus of the receptor mouse for gestation, thus obtaining the positive offspring transgenic mouse; and mating the positive offspring transgenic mouse with a transgenic mouse with specific expression of tTA or a transgenic mouse withspecific expression of rtTA, and picking out the inducible transgenic mouse cardiomyopathy animal model from the hybrid offsprings. The invention further discloses applications of the transgenic mousecardiomyopathy animal model obtained through the construction method in screening medicines for treating cardiomyopathy. According to the technical scheme provided by the invention, the modeling success rate is high, the phenotype is stable, the cost is ow, and whether inducting for the generation of cardiomyopathy is needed or not can be selected according to the actual demands.

The invention relates to a construction method of an STAT3 mitochondrial localization conditional gene knock-in mouse model. The method comprises the following steps: firstly, constructing an STAT3 mitochondrial localization conditional gene knock-in recombinant targeting vector, transfecting the recombinant targeting vector to fertilized ova of a donor mouse, and then transplanting the donor mouse fertilized ova into the uterus of a fake pregnant mother mouse to produce a chimeric mouse; mating the chimeric mouse with a C57BL / 6J mouse to obtain an F1 generation; mating the F1 generation witha mouse of tissue-specific expression Cre recombinase to obtain a double-hybrid mouse; and are intercrossing the double-hybrid mouse and an STAT3<f / +> or STAT3<f / f> mouse, and inducing the obtained offspring by tamoxifen to obtain the STAT3 mitochondrial localization conditional gene knock-in mouse model. According to the mouse model disclosed by the invention, the STAT3 gene is expressed in specific tissues and cell types, and an ideal model is provided for researching the action mechanism of STAT3 in specific cell development and differentiation, body metabolism and tumor occurrence and development.

The invention relates to the technical field of medicines, and provides applications of uterine cavity fluid-derived exosome in preparation of drugs and adjuvant therapeutic agents for treating infertility related diseases. Through the verification of experiments, exosome realizes the treatment or relieving of endometrial growth disorders by promoting the increasing of the thickness of endometrium, enhancing the activity of endometrial cells, reducing the damages to intimal cells, promoting VEGF expression, maintaining the dryness of endometrial stromal cells and stimulating proliferation; andthrough the realizing of the treating or relieving of maternal-fetal immune tolerance disorder-related diseases by achieving immune tolerance effects, treating spontaneous abortion and enhancing T helper lymphocyte levels, a novel theoretical basis can be provided for infertility treatment. In addition, for both fertilized eggs being fertilized in vivo or fertilized in vitro, the exosome can effectively enhance developmental rates in multiples; and implantation rates and farrowing rates of in vitro fertilization transplantation can be increased as well, so that positive auxiliary effects on infertility treatment can be achieved.

The invention belongs to the field of biotechnology, and relates to a resource library of rat strains with gene mutation and a preparation method thereof. The rats with gene mutation in the resource library can be obtained from rat spermatogonial stem cells, embryonic stem cells or fertilized ovum with piggyBac transposon insertion mutation. The rats with gene mutation in the resource library also can be obtained from shifting the transposon in germ cells of male rats with mutation. The library can be applied to animal phenotype analysis, gene function research, human disease simulation, medicament screening and drug target development.

The invention discloses a high-survival-rate sheep oosperm culture method for Hu sheep embryo in-vitro culture. The high-survival-rate sheep oosperm culture method comprises the following steps: (1) oocyte collection: sucking oocytes from ovaries of Hu sheep; (2) sperm collection: extracting sperms from the male Hu sheep; (3) fertilization: carrying out artificial fertilization on eggs and sperms;(4) embryo culture: putting the fertilized eggs into an incubator for culture; and (5) observation: observing the development condition of the embryo, wherein the incubator in the step (4) comprisesa main body, a feeding device, a rotating frame, a culture device and a discharging device, wherein the feeding device comprises a feeding channel, a conveying mechanism, a feeding opening, a rotatingtable and a transition mechanism. According to the method, the embryo survival rate is increased through artificial fertilization, the transition mechanism isolates the embryos from outside air whenthe embryos are placed into the incubator, so that embryo development is prevented from being influenced, and the discharging device realizes the purpose of independent taking a culture dish, so thatthe survival rate of embryos is increased.

Provided are a method for preparing a mammalian ovum or embryo in which zona pellucida has been thinned or eliminated, and a method for fertilization using the mammalian ovum prepared by the aforementioned method. The method for thinning or eliminating zona pellucida of a mammalian ovum or embryo involves treating or culturing the mammalian ovum or embryo (such as an unfertilized ovum, a fertilized ovum, or an embryo in the early stages of development) in a culture medium containing a reducing agent having SH groups (such as reduced glutathione or DTT). The resulting mammalian ovum or embryo (such as an unfertilized ovum, a fertilized ovum, or an embryo in the early stages of development) in which zona pellucida has been thinned or eliminated is capable of realizing an improved fertilization rate and development rate when used for in vitro fertilization, transplantation of a fertilized ovum, or preparation of an embryo in the early stages of development used in the production of a genetically modified animal.

The invention discloses an animal sex control method based on Rbmy gene editing and application. The method controls animal sex by using CRISPR / Cas9 system to specifically cut Rbmy gene on Y chromosome. Rbmy gene encodes a germ cell specific nucleoprotein, this protein is an important factor related to spermatogenesis, and by cutting Rbmy gene, it is possible to deactivate Rbmy gene, thus Y sperm or fertilized ovum with Y sperm is deactivated and cannot develop into a normal embryo, birth ratio of female animals is finally increased and sex control is achieved. Therefore, sex ratio of animal offspring can be controlled through the method so that the ratio of female animals with better economic value is increased and production efficiency and economic effect are greatly improved.

The invention provides a method for obtaining a sterile mammal with rapidness and high efficiency. The method comprises the following steps: a) obtaining a sperm of a male mammal, and subjecting the obtained sperm to capacitation; b) obtaining an ovum of a female mammal; c) preparing an oosperm by utilizing the sperm obtained in the step a) and the ovum obtained in the step b); d) obtaining an aspermous male mammal; e) preparing a pseudopregnant female mammal; f) preparing an embryo by utilizing the oosperm obtained in the step c); g) obtaining a pseudopregnant female mammal containing the embryo obtained in the step f); and h) subjecting the female mammal obtained in the step g) to sterile culture so as to obtain the sterile mammal. The method provided by the invention has the advantages of short time, fast action, high efficiency and small breeding population, so the sterile mammal used for various purposes can be provided with rapidness and high efficiency.

The invention discloses an automatic IVF culture and time difference analysis device and a matched culture dish. The device is composed of a main device, a matched gas-liquid flow device and an integrated controller; and the main device comprises a heat insulation sealing cover, a sample adding device, a culture dish bearing device, and a CO2 concentration, O2 concentration, humidity and temperature monitoring and adjusting device. The culture dish is provided with an oil seal cavity groove, a small culture chamber, a culture dish culture solution feeding hole and a culture dish waste discharge hole, and a culture solution buffer cavity communicated with the culture dish culture solution feeding hole and the injection port of the small culture chamber and a waste liquid buffer cavity communicated with the culture dish waste discharge hole and the waste discharge hole of the small culture chamber are arranged below the oil seal cavity groove. Relative constancy of a culture environmentcan be maintained, a spiral flow liquid changing technology can guarantee the removing effect of sperm cells, cumulus cells and metabolic waste, concentration gradient changing of all the culture solutions is achieved, external stimulation to fertilized eggs is reduced to the maximum extent, and the fertilization rate and the blastocyst cultivation rate are effectively increased.

The disclosed invention comprises a device that utilizes a manually controlled articulating arm to make a precise, small abrasion on the intrauterine wall prior to ovulation. The preferred embodiment of the invention comprises a handle, an arm having a rigid portion and an articulating tip both of which are covered by a casing having non-irritant properties, wherein the handle is connected to the arm by a connection member that contains a trigger mechanism operable to move the articulating tip by pulling a trigger.

The invention discloses a method for carrying out ICSI (intracytoplasmic sperm injection) on a pig. The method for carrying out ICSI on the pig comprises the following steps: (1) hydrolyzing sperms in vitro animal semen with lipoidase or lipoidase solution, and collecting a hydrolysis product, so that the sperms used for preparing ICSI fertilized ova; (2) injecting the sperms used for preparing the ICSI fertilized ova into in vitro oocytes to obtain ICSI ova; and (3) carrying out electric activation on the ICSI ova and cultivating, so that the ICSI fertilized ova are obtained. The method for carrying out ICSI on the pig has the advantages that bad effects on depolymerization of pig sperms, formation of male pronucleus and later development due to addition of PVP can be avoided, and regions such as plasma membranes and perforatorium of the pig sperms can not be damaged, so that release of oocyte activation promoting factors from the pig subjected to ICSI operation can be facilitated, and the formation rate of the male pronucleus and the formation rate of blastocysts of embryos after fertilization are increased.

A livestock fertilized-egg recovery device (10) is provide with: a perfusate supply part; a first syringe (20) for suctioning a perfusate from the perfusate supply part and injecting the perfusate into the uterus of livestock; a balloon catheter for injecting the perfusate into a uterine horn and recovering the perfusate together with a fertilized egg; a second syringe (30) for recovering the perfusate injected into the uterus of livestock and discharging the perfusate into a fertilized-egg recovery part; said fertilized-egg recovery part; liquid delivery lines (211, 212, 311, 312) that respectively connect between the perfusate supply part, the first syringe, the balloon catheter, the second syringe, and the fertilized-egg recovery part; and a liquid delivery direction control part (50) for controlling the delivery direction of the perfusate within the liquid delivery lines, wherein the livestock fertilized-egg recovery device is also provided with syringe pump control parts (40, 40')for controlling the pumping action of the two syringes.

The invention relates to a retrievable intrauterine device which includes a recess (10) suitable for containing one or more elements selected from the group that includes an embryo, male and / or femalegametes, a fertilised oocyte, a non-fertilised ovule and a combination of said elements. The recess (10) includes an inner wall (11) made of a biocompatible polymer, the inner wall (11) including atleast one perforated portion (13a, 13b) having perforations with dimensions of between 1 [mu]m and 10 [mu]m, and an outer wall (12) covering the inner wall (11), the outer wall (12) being made of a biocompatible silicone elastomer and including at least one opening (14a, 14b) opposite said at least one perforated portion (13a, 13b) of the inner wall (11). The use of the device relates to the pre-implantation development of gametes and / or embryos.

The invention aims to diagnose pregnancy in a livestock animal by a method which can be carried out at a breeding site while minimizing invasion of the subject animal and fetuses thereof. The method according to the present invention for diagnosing pregnancy in a ruminant comprises: a measurement step of using a sample containing mucosal epithelial cells collected from the genitals ranging from the uterine cervical canal to the vulvar orifice of the subject ruminant, said subject ruminant being in the state corresponding to the early stage of pregnancy, and measuring the expression level of amolecule, which is induced in response to IFN-[tau], in the mucosal epithelial cells; and a diagnosis step for comparing the expression level with an expression level of the aforesaid molecule in corresponding mucosal epithelial cells of a non pregnant ruminant of the same species and thus diagnosing pregnancy in the subject ruminant. According to the present invention, pregnancy can be quickly diagnosed before a new ovulation cycle starts after breeding, artificial insemination or fertilized egg transplantation.

A recoverable intra-uterine device comprises a housing (11) containing one or several elements selected from a group containing an embryo, male and / or female gametes, a fertilised oocyte, an unfertilised egg and the combination thereof, wherein said housing (11) is provided with a wall made of a biocompatible material. The wall is provided with a series of perforations (12, 12') whose size is sufficient in order to bring the intra-uterine medium into a cellular contact with the housing (11) and to keep the elements therein. The inventive device is provided with a system for loading and unloding one or several elements selected from the group containing an embryo, male and / or female gametes, a fertilised oocyte, an unfertilised egg and the combination thereof.

The invention discloses a composite additive for improving reproductive performance of female raccoon dogs. The composite additive is fed in a mating period, can enhance ovarian function, promote follicle development and maturation, effectively reduce the phenomena of estrus retardation, non-concentration, unconspicuousness, no ovulation, missed mating and the like of female raccoon dogs, and improve the mating rate and reproductive rate of female raccoon dogs. After mating, the additive is fed, luteinizing in bodies of the female animals is promoted, implantation of fertilized eggs is promoted, the phenomena that the fertilized eggs are not implanted, false pregnancy occurs, nonpregnant occurs, embryonic absorption occurs and the like are prevented and reduced, and the pregnancy rate andthe farrowing rate of the female animals are increased; the late pregnancy period is the highest nutritional requirement period of the fetus, and the product can promote the nutritional absorption ofthe fetus and ensure the healthy development of the fetus, thereby reducing the occurrence of dead fetus, rotten fetus, dystocia, abortion, weak cubs and the like; and when the product is continuouslyused in the lactation period, female animals can be promoted to secrete milk, the lactation yield is increased, the lactation period is prolonged, and the phenomena of weak piglets, turning around and the like caused by hypogalactia are reduced, so that the survival rate, the healthy piglet rate and the uniformity of the piglets are improved.

The invention relates to an expression vector containing a calpain-1 gene. The expression vector comprises a Tet operon, an alpha-MHC promoter nucleotide sequence, a mitochondrial signal peptide sequence and a gene segment of calpain-1 which are connected in sequence. The invention further provides a construction method of an inducible transgenic mouse cardiomyopathy animal model. The constructionmethod comprises the following steps: importing the expression vector containing the calpain-1 gene into oosperms of a mouse, and transferring the oosperms into the uterus of the receptor mouse for gestation, thus obtaining the positive offspring transgenic mouse; and mating the positive offspring transgenic mouse with a transgenic mouse with specific expression of tTA or a transgenic mouse withspecific expression of rtTA, and picking out the inducible transgenic mouse cardiomyopathy animal model from the hybrid offsprings. The invention further discloses applications of the transgenic mousecardiomyopathy animal model obtained through the construction method in screening medicines for treating cardiomyopathy. According to the technical scheme provided by the invention, the modeling success rate is high, the phenotype is stable, the cost is ow, and whether inducting for the generation of cardiomyopathy is needed or not can be selected according to the actual demands.

The invention discloses a combined culture dish for assisted reproduction. The combined culture dish comprises a first culture dish body and a second culture dish body, wherein an ovum containing groove is fixedly installed in the middle of the bottom in the first culture dish body, a plurality of sperm containing grooves are fixedly installed in the periphery of the bottom in the first culture dish body, the second culture dish body is movably installed at the upper part in the first culture dish body and is arranged on the upper sides of the ovum containing grooves and the sperm containing grooves, a normal embryo culture groove is fixedly installed in the middle of the bottom in the second culture dish body, and a special embryo culture groove, a frozen embryo culture groove and a transplanted embryo culture groove are fixedly installed in the periphery of the bottom in the second culture dish body. According to the combined culture dish for assisted reproduction, the first culturedish body and the second culture dish body can be movably installed together, high-quality fertilized ova are screened in the first culture dish body and then are separately cultured in the second culture dish body, the frequency of exposing the culture dishes in an incubator can be reduced, errors are reduced, and embryos in the culture dishes are effectively protected.

Described herein are methods for synchronizing ovulation in a herd, inducing superovulation in a single animal, and improving frequency of successful implantation and development of fertilized ova.

A recoverable intra-uterine device comprising a housing containing one or several elements selected from a group containing an embryo, male and / or female gametes, a fertilised oocyte, an unfertilised egg and the combination thereof, wherein said housing is provided with a wall made of a biocompatible material. The wall is provided with a series of perforations whose size is sufficient in order to bring the intra-uterine medium into a cellular contact with the housing and to keep the elements therein. The inventive device is provided with a system for loading and unloading one or several elements selected from the group containing an embryo, male and / or female gametes, a fertilised oocyte, an unfertilised egg and the combination thereof.

The invention aims to provide an artificial insemination and oosperm hatching technology of whitebait in severe cold regions, wherein the process includes an artificial insemination process and an oosperm hatching process. The quality of produced bred fries is high, and the quality is high.

The invention relates to a method and application for establishing an Arvcf gene knockout mouse model, wherein the mouse model is a mouse that knocks out seven exons in the Arvcf gene, and discloses a method for establishing an Arvcf gene knockout mouse model. method of the mouse model. First determine the knockout region of the gene to be knocked out Arvcf, then design and synthesize specific sgRNAs and corresponding primers, then linearize and purify the constructed sgRNA / Cas9 vector for in vitro transcription, and finally inject together by microinjection The fertilized eggs of mice were transplanted into the uterus of pseudopregnant female mice to establish the Arvcf gene knockout mouse model. The tails of the obtained progeny mice were collected and DNA was extracted, and the obtained mice were confirmed to be wild-type or homozygous or heterozygous for the target region of the Arvcf gene by using two pairs of primers through two PCR reactions and sequencing. mice. This knockout mouse model can be used to study the role of ARVCF in the psychiatric diseases involved.

The invention discloses a pipette egg transfer needle adapter. The pipette egg transfer needle adapter comprises a spearhead interface, a fastening nut and a sealing rubber ring group; the spearhead interface, the fastening nut and the sealing rubber ring group are all of revolving body structures, through holes are formed in the spearhead interface, the fastening nut and the sealing rubber ring group in the axial direction, the fastening nut sleeves the front of the spearhead interface, and the fastening nut and the spearhead interface are connected by threads. The sealing rubber ring group is installed in an inner cavity of the fastening nut and is pressed between the spearhead interface and the fastening nut. The pipette egg transfer needle adapter has the beneficial effects that the adapter can form a simple and convenient pipette egg transfer needle system with various kinds of egg transfer needles, compared with an original method, the processing and transfer of various embryos such as follicle removal, oocyte, fertilized eggs, blastaea can be more easily, quickly, and accurately realized, granular cells are separated, the activity of the embryos during operation is ensured,and efficiency and reliability of embryo processing and transfer operation are improved.