178 results about "Embryo culture" patented technology

The invention discloses a pig chimeric embryo culture medium and application thereof in effectively improving the in-vitro chimeric ability of pig embryonic stem cells, and belongs to the technical field of bioengineering. According to the pig chimeric embryo culture medium disclosed by the invention, a neutral culture environment for promoting synchronous development of stem cells and embryos is created by accurately regulating and controlling growth factors, nutrient substances and other key components in the culture medium. On the basis, overexpression of anti-apoptosis factors, optimization of microinjection conditions and adoption of a laser-assisted incubation means are combined, so that the chimeric efficiency is further improved, integration of the stem cells and embryos is effectively supported, the chimeric efficiency is improved, and a reliable in-vitro tool is provided for evaluating the chimeric ability and pluripotency of the pig stem cells.

The invention relates to the technical field of embryo culture platforms, in particular to a microfluidic-AI monitoring-metabolism regulation embryo culture platform and a preparation method thereof.The microfluidic-AI monitoring-metabolism regulation embryo culture platform comprises a microfluidic culture system used for achieving embryo culture, metabolite removal and culture medium replacement; the artificial intelligence monitoring system is used for remotely monitoring the embryonic development state in real time and predicting the embryonic development condition; the metabolism regulation and control system is used for feeding back and optimizing culture conditions online according to an artificial intelligence monitoring result and a metabolite analysis result; the intelligent control module is electrically connected with the micro-fluidic culture system, the artificial intelligence monitoring system and the metabolism regulation and control system, and the micro-fluidic culture system can accurately control the culture environment on the micrometer scale, including temperature, pH value and gas composition parameters. The dynamic fluid environment is closer to the in-vivo physiological environment of the embryo. The system can simulate periodic nutrient supply and metabolic waste removal in the uterus, and provides optimal growth conditions for embryos.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a device and a method for gradually adjusting the glucose concentration of an in-vitro embryo culture solution. The device comprises at least two miniature liquid storage bins, wherein each miniature liquid storage bin is filled with a high-concentration glucose solution in advance; the slow release channel is respectively connected with each micro liquid storage bin and the culture solution main container; the culture solution main container is used for accommodating an embryo and a basal culture medium; the control unit is used for sequentially opening the release ports of the miniature liquid storage bins, so that the high-concentration glucose solution sequentially flows into the culture solution main container through the slow release channels. The technical means of staged closed-loop micro sugar control is introduced into the structure and the using method of the device, so that the accurate adjustment of the sugar concentration of the in-vitro embryo culture solution is realized, the requirements of embryos in different development stages can be effectively met, and the embryo culture quality and the subsequent development potential are remarkably improved.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The invention discloses an embryo culture method and an embryo culture solution for improving the quality of a pig somatic cell nuclear transfer embryo, and a small molecule substance X1 capable of specifically binding to a RepA structural domain of Xist is added into a pig early embryo culture solution, so that the development quality of a pig SCNT embryo is improved.

The invention relates to the technical field of in-vitro embryo culture, in particular to an in-vitro culture method and culture system of a mouse embryo. The method comprises the following steps: dropwise adding hydrogel onto a PDMS substrate, putting at least one, at least two, at least three, at least five or more embryos into hydrogel liquid drops, curing, and then adding a culture solution for embryo culture; wherein the Young modulus of the cured hydrogel liquid drops is 10 to 100 Pa. The in-vitro culture system comprises a PDMS (Polydimethylsiloxane) substrate and hydrogel liquid drops arranged on the PDMS substrate, and the hydrogel liquid drops are used for coating the mouse embryo; wherein the Young modulus of the cured hydrogel liquid drops is 10 to 100 Pa. When the method and the system are used for in-vitro embryo culture of mice, the embryos can normally develop and have a relatively high development rate within the in-vitro culture time of 9 days during in-vitro embryo culture.

The invention discloses a biomarker combination and application thereof in predicting embryo quality. Wherein the biomarker combination comprises small molecule non-coding RNA (Ribonucleic Acid) with nucleotide sequences as shown in SEQ ID NO.1-3; or micromolecule non-coding RNA as shown in SEQ ID NO.1-8; or micromolecular non-coding RNA as shown in SEQ ID NO.1-13. The invention also discloses a method for preparing the micromolecular non-coding The biomarker combination disclosed by the invention not only contains miRNA, but also contains multiple types of sncRNA such as tsRNA, snRNA, Y RNA, snoRNA and the like, and the biomarker combination has the characteristic of obviously distinguishing the embryo quality. When the biomarker combination is used for predicting the embryo quality, only an embryo culture solution is needed, and as a non-invasive sample collection operation, the shape and subsequent development of the embryo are not influenced. Meanwhile, the invention also provides a construction method and a prediction method of the embryo quality prediction model, and the final model has high sensitivity and high specificity, thereby providing guidance for embryo screening.

The invention discloses a technology for shortening a cotton breeding cycle and application thereof, and the technology comprises the following steps: 1, accelerating budding and flowering under mixed light of blue light, green light, red light and far-red light; 2, performing GA3 treatment to promote germination of immature embryos; 3, using the created L-mAUH and mAUH cotton immature embryo culture medium to culture the 25-28-day-old cotton immature embryos into seedlings; fourthly, the first step, the second step, the third step and the fourth step are circularly carried out for multiple times, in the process, molecular markers are used for carrying out auxiliary selection on the good characters of the foreground, and meanwhile whole genome re-sequencing is used for carrying out background auxiliary selection. The method has the advantages that by means of efficient blue, green, red and far-red spectrum adjustment, GA3 treatment, immature embryo culture, molecular marker selection and whole genome re-sequencing, cotton growth and development can be remarkably accelerated, and the cotton breeding period is effectively shortened. According to the method, a conventional backcross breeding process can be effectively integrated, excellent characters can be quickly transformed, and an improved line with a high genetic background recovery rate can be obtained within 1.5 years.

The invention discloses a precise clamping type multi-station embryo culture dish operation table and a use method thereof, and relates to the technical field of embryos. A guide groove is formed in the inner side of a guide frame, a plurality of alignment flat grooves are formed in the top side face of the guide frame, and a plurality of spiral fasteners aligned with the alignment flat grooves are installed on the side face of the guide frame in a screwed mode. The clamping tooth piece comprises a toothed plate, two vertical plates fixed to the upper side of the side end of the toothed plate and a transverse rod fixed between the upper side ends of the two vertical plates, and a sliding piece installed at the guide groove in a sliding mode is further fixed to the side end of the toothed plate. The embryo culture dish comprises a lower ring body and an upper ring body, and a ring gap is formed between the periphery of the lower ring body and the upper part of the upper ring body. The gear ring piece comprises a ring plate which is positioned and sleeved at the position of the ring position notch; and an outer ring tooth opening matched with the side tooth opening is formed in the outer ring side of the ring plate. The culture dish is effectively prevented from being mistakenly touched and shaken, the problem that normal development of cells is affected by a high-intensity magnetic field and the like is also avoided, and a stable external environment is provided for embryonic cell culture.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a glucose slow-release structure used in an embryo in-vitro culture solution, a preparation process and a use method. The glucose slow-release structure comprises at least one inner layer containing glucose and a biodegradable material outer layer wrapping the inner layer, and the biodegradable material outer layer comprises a matrix with a plurality of micropores and at least one sealing material layer used for sealing pores of the micropores. According to the invention, gradient release from low sugar in the cleavage stage to high sugar in the blastocyst stage is realized, midway liquid change or artificial sugar supplement is not needed, the operation is obviously simplified, and pollution and mechanical stress risks are reduced.

The invention relates to an artificial intelligence prediction method and system for a multi-embryo blastocyst culture outcome. The method comprises the following steps: acquiring morphological parameter data of embryo culture; constructing a feature vector based on the morphological parameter data; and inputting the feature vector into a preset classifier, and predicting a culture result of the embryo group. The method is suitable for various culture environments, and not only can treat group culture, but also can be applied to period prediction of individual culture. The diversity meets the gap among different culture strategies in the prior art; while the dependence on individual labels is reduced, the prediction accuracy of the method is equivalent to that of an existing method based on single embryo probability prediction, and the technical reliability is ensured.

The invention relates to an anti-browning method in a coconut microspore somatic embryo culture process, and belongs to the technical field of coconut tissue culture. The anti-browning method in the coconut microspore somatic embryo culture process comprises the following steps: S1, anther collection and pretreatment; s2, anther observation, disinfection and separation; s3, anther culture and embryo culture. According to the anti-browning method in the coconut microspore somatic embryo culture process, PVP is added into the culture medium to resist the browning phenomenon in the coconut microspore embryo induction process; when the explant is cut, a citric acid solution is dipped to reduce browning caused by physical damage; ascorbic acid (vitamin C) is also added into the culture medium to adjust the pH value and resist the browning phenomenon; in addition, low-concentration TDZ (thidiazuron) is used as an induction hormone, so that the embryo induction efficiency is improved.

The invention provides an in-vitro embryo culture solution and a preparation method thereof.The IVM solution of the in-vitro embryo culture solution comprises CaCl2, KCl, KH2PO4, glycine, HEPES sodium salt, lactic acid, MgSO4, NaHCO3, inositol, glucose, essential amino acid, non-essential amino acid, retinoic acid, taurine and hormones, and when the in-vitro embryo culture solution is used as mammals such as cows, the in-vitro embryo culture solution can be used for preparing the in-vitro embryo culture solution. And the IVF embryo production efficiency and embryo quality of the cattle can be improved.

The application discloses an embryo culture method and embryo culture solution for improving the quality of pig somatic cell nuclear transfer embryos, wherein a small molecule substance X1 capable of specifically combining with the RepA domain of Xist is added into a pig early embryo culture solution, so as to improve the development quality of pig SCNT embryos.

The invention belongs to the technical field of bioengineering, and particularly relates to application of RPS26 recombinant protein in promoting early embryonic development of animals. Experiments show that after the RPS26 recombinant protein is added in the early-stage in-vitro embryo culture process of the lamb goats, the blastocyst rate of the lamb goats can be increased to 40.0%; meanwhile, the cell proportion of cell mass in the blastocyst can be remarkably increased, the cell proportion of trophoderm is reduced, and the total number of blastocyst cells is not influenced, so that the blastocyst development rate can be remarkably increased and the blastocyst quality can be improved by utilizing the RPS26 recombinant protein.

The invention discloses a method for determining a chromosome copy number and a specific gene carrying chain by using an embryo culture solution. The method can be used for simultaneously detecting embryo chromosome abnormality and a specific gene carrying state. The method comprises the following steps: firstly, collecting blood sample DNA of two specific gene carriers, a pre-certified person or a last generation of relatives; constructing an STR haplotype, designing an STR site primer for amplification, and determining a family haplotype through electrophoretic analysis; the method comprises the following steps: carrying out whole genome amplification on a collected trace free DNA sample of a trace cell or embryo culture solution by using a Pi coPLEX method, respectively designing primers for amplification products aiming at monogenic disease types and mutation sites, and directly detecting pathogenic genes; performing second-generation sequencing, and comparing with a normal sequence to analyze and judge the copy number of chromosomes; primer amplification is designed according to the constructed family distinguishing type STR site, STR site haplotype information is obtained, whether a specific gene variation chain is inherited or not is judged, and therefore an euploid embryo which does not carry a specific gene is efficiently screened out.

The invention provides a method for constructing an organ regeneration genetic compensation effect, and relates to the technical field of biology. Comprising the following steps: S1, designing sgRNA; s2, carrying out in-vitro transcription and purification to obtain a high-purity sgRNA solution; s3, preparing fertilized eggs, preparing sgRNA mixed injection, and performing embryo culture after micro-injection operation to obtain F0-generation animal larvae; s4, performing gene knockout efficiency detection on the F0-generation animal larvae, screening out the F0-generation animal larvae with high knockout efficiency, and breeding and mating the F0-generation animal larvae to obtain F1-generation animals; and S5, verifying and screening the F1-generation animals to obtain homozygous knockout individuals, carrying out amputation treatment on the homozygous knockout individuals, and verifying the genetic compensation effect of organ regeneration. The YAP knockout animal is constructed through a gene editing technology, the effect and mechanism of the YAP knockout animal in limb regeneration are researched, whether YAP deletion affects the regeneration capacity through a genetic compensation effect or not can be revealed, and then a theoretical basis and an experimental basis are provided for future regenerative medicine application.

The invention provides an early-maturing peach hybrid seed pretreatment method and application thereof in direct sowing and seedling raising, and belongs to the technical field of plant breeding. The invention provides a pretreatment method of early-maturing peach hybrid seeds, which comprises the following steps: disinfecting completely-matured early-maturing peach hybrid fruits, taking out seed kernels, mixing the seed kernels with a culture solution, carrying out ultrasonic treatment, and drying in the shade. By utilizing the pretreatment method disclosed by the invention, the seeds which cannot completely develop embryos due to the short development period of fruits, finally have no direct seeding breeding prospect and even are not suitable for embryo rescue can be subjected to multiple stimulation by multiple nutrient substances and plant growth regulators in the culture solution under the microwave condition, so that the seeds can be fully cultured. The seed kernels continue to grow and develop after being in vitro, and the wizened or gelatinous seeds without embryo culture are subjected to in-vitro development and after-ripening. According to the seeds treated through the pretreatment method, dormancy is almost broken, the low-temperature stratification time can be shortened, and even direct seeding breeding can be achieved without low-temperature stratification treatment.

This invention discloses a culture medium additive for improving embryonic developmental potential and its application, belonging to the field of in vitro embryo culture technology. This invention is the first to discover that CPI-455, a KDM5 histone demethylase inhibitor, can significantly improve the blastocyst rate, developmental speed, and embryo quality of preimplantation embryos by increasing H3K4me3 levels within a certain concentration range. Simultaneously, the addition of CPI-455 and NMN can substantially increase the rate of high-quality / high-quality blastocysts and embryo quality in in vitro embryo culture. The embryonic developmental potential additive provided by this invention is of great significance for improving in vitro embryo culture efficiency and embryo quality, accelerating the process of breeding superior embryos, and assisting in research on embryo-related mechanisms.

The present invention belongs to the field of in vitro embryo technology and relates to the technology of in vitro cultivation of pre-implantation embryos of mammals. The present invention provides the use of dihydromyricetin or its derivatives in the preparation of products for improving the quality and developmental ability of in vitro embryos, and products for improving the developmental ability of in vitro embryos, the use of dihydromyricetin or its derivatives in improving the quality of in vitro embryos, and a method for improving the developmental ability of in vitro embryos. The present invention discovers that dihydromyricetin or its derivatives can increase the blastocyst rate of in vitro embryo culture, reduce the content of cellular reactive oxygen species, increase the content of cellular glutathione, and promote the expression of oxidative stress genes, thereby effectively improving the quality and developmental ability of in vitro embryos.

The utility model relates to the technical field of portable embryo incubators and discloses a portable embryo incubator which comprises an incubator body, an incubator cover arranged on the upper side of the incubator body and a control panel arranged on one side of the incubator body, a uniform thermal circulation mechanism is arranged in the incubator body, and a stable transfer mechanism is arranged on the lower side of the incubator body. The uniform heat circulation mechanism comprises a heat flow cavity, the heat flow cavity is formed in the cultivation box, a double-shaft motor is fixedly connected to the interior of the cultivation box, drainage fan blades are fixedly connected to the output end of the double-shaft motor, and the uniform heat circulation mechanism is used for providing uniform heat flow for test tubes; according to the mechanism, the sealing performance of the interior of the incubator can be improved, meanwhile, hot air flow can be uniformly dispersed, the stable transfer mechanism is used for providing convenient transfer for the incubator, test tubes are convenient to be perpendicular to the ground all the time, and meanwhile, the mechanism enables the incubator to be relatively stable during angle adjustment through an elastic structure and air pressure.

The invention provides a method for simultaneously detecting various water-soluble vitamins in a human assisted reproduction culture solution, which can simultaneously detect at least four vitamins, including vitamin B1, vitamin B2, vitamin B5 and vitamin B6. According to the method, the content of the water-soluble vitamins in the human assisted reproduction culture solution can be effectively monitored, and the safety of the embryo culture process is ensured.

The invention provides a wide-compatibility sweet persimmon rootstock Taifu tissue culture and rapid propagation method, and belongs to the technical field of plant tissue culture. According to the method, young fruits pollinated for 58-62 days by Taifu are selected, young embryos of the young fruits are taken as explants and inoculated into a culture medium to be cultured, the culture medium comprises the following components with the concentration: (1 / 2N) MS, 0.07-0.09 mg.L <-1 > IBA, 4.8-5.2 mg.L <-1 > ZT, 70-80 mg.L <-1 > cane sugar, 7-9 g.L <-1 > agar and 480-520 mg.L <-1 > PVP, the pH value is 5.8-6.0, after embryo culture seedlings are obtained, root cutting treatment is conducted, then dark culture is conducted for 5-6 days, and Taifu tissue culture seedlings are obtained. According to the method, the concentration of each component in the culture medium is adjusted, and root promotion is carried out by means of root cutting treatment and dark culture, so that browning of embryo culture seedlings is avoided, tissue culture seedlings with good growth vigor are obtained, and the survival rate after transplanting is improved.

The utility model provides a multifunctional embryo culture dish for reproductive medicine, which belongs to the field of reproductive medicine and comprises a dish bottom, a central well is arranged at the top of the dish bottom, a waste liquid well is arranged on one side of the central well, a culture hole is arranged on one side of the waste liquid well, a placing groove is arranged in the dish bottom and is positioned right below the culture hole, and the culture hole is arranged in the placing groove. And a temperature control base is arranged in the placement groove. Through the arrangement of the central well, the waste liquid well, the culture holes, the placing groove and the temperature control base, the waste liquid well, the culture holes and the temperature control base can be additionally designed under the condition that the central well is reserved, the waste liquid well can be used for storing waste liquid, and the culture holes are multiple, so that more than two embryos can be cultured; various experimental data can be conveniently compared, the temperature control base can keep the temperature in the culture hole, the temperature is constant at about 37 DEG C, and the survival rate of embryos is increased.

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 utility model discloses a movable embryo laboratory which comprises a movable chassis and a laboratory box body, the laboratory box body is arranged above the movable chassis, and a traction device is fixedly arranged at the front end of the walking direction of the movable chassis. A unit operation and maintenance room, a disinfection room, an ovum collection room, an in-vitro fertilization culture room and a cryopreservation room are arranged in the laboratory box body from front to back in the walking direction of the movable chassis, and every two adjacent areas of the ovum collection room, the in-vitro fertilization culture room and the cryopreservation room are communicated through doors. Wall bodies of the disinfection room, the ovum collection room, the in-vitro fertilization culture room and the cryopreservation room are provided with delivery windows for communicating two adjacent areas, and the disinfection room is partitioned in the front-back direction to form a gas cylinder room and a decontamination room. According to the utility model, sperm and egg sample treatment time can be shortened, and embryo culture success rate is improved; sample loss caused by long-distance transportation is avoided; the operation flexibility is improved, and the embryo treatment cost is reduced; the method is convenient to deploy quickly and is suitable for pastures under different terrains and climate conditions.

The invention relates to a culture medium for constructing a single-cell-initiated protointestinal-like embryo to an organoid embryogenesis stage embryo, the culture medium comprises an AL culture medium and a PrE induction culture medium, the AL culture medium is formed by adding LY2090314 and AS1842856 into an N2B27 basic culture medium, the LY2090314 is 10nM, the AS1842856 is 0.6 mu M, the PrE induction culture medium is formed by adding a pharmaceutical composition into the N2B27 culture medium, and the ALE culture medium is a PrE induction culture medium. The pharmaceutical composition is composed of 2.5 [mu] M of 1-Azakenpaullone, 0.2 [mu] M of TTNPB, 100 ng / ml of FGF4 (Fibroblast Growth Factor 4) and 1 mM of 8Br-cAMP. The invention also provides an application of the embryo culture medium and a construction method from a protointestinal-like embryo starting from a single cell to an organoid embryogenesis stage embryo. According to the method, a continuous construction scheme from the protointestinal-like embryo starting from a single cell to the organoid embryogenesis period embryo is adopted, so that the construction mode is simplified, the construction efficiency is improved, the application value of the embryoid is enhanced, and a good platform is provided for embryonic development research and disease drug screening.

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.

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.

This invention provides an independent embryo culture dish, comprising a dish body, several culture chambers, and several waste liquid tanks. The culture chambers are arranged in a matrix on the dish body and connected to the dish body. A waste liquid tank is provided on one side of each row of culture chambers. The interior of each culture chamber is divided into an embryo culture layer, a covering oil layer, and an overflow control layer from bottom to top. A through-channel is provided between the overflow control layer and the waste liquid tank, and the overflow control layer and the waste liquid tank are connected by the through-channel. By setting an embryo culture layer, a covering oil layer, and an overflow control layer arranged from bottom to top in each independent culture chamber, this invention not only effectively ensures a stable environment for the embryo culture medium, but also achieves automatic discharge of excess culture oil through the through-channel structure between the overflow control layer and the waste liquid tank, avoiding culture environment disorder and cross-contamination caused by excessive oil layer thickness.