122 results about "Embryo culture" patented technology

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 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 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 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 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.

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 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.

The invention discloses a method for improving the normal fertilization rate of an in-vitro ICSI embryo of a dairy cow and reducing the abortion rate, which comprises the following steps: preparing an ICSI egg from X-sex controlled sperm and an oocyte which is cultured by a mature oocyte and is used as a donor by adopting a microinjection mode, the ICSI eggs are incubated in an oocyte maturation culture solution (OM) for 3-5 h and then transferred to ETH to be activated for 4-6 min so as to promote normal fertilization of the ICSI eggs, then embryo culture is conducted, and then sexual control embryos are obtained; the ETH refers to an oocyte maturation culture solution containing 6 to 8 percent of ethanol. The normal fertilization rate of the ICSI embryos produced by the method is remarkably improved, and the early abortion rate is remarkably reduced in inspection after transplantation.

The application provides an in-vitro seed embryo culture method of peony hybrid seeds, which comprises the following steps: 1) taking Chuan-shao No.1 or Chuan-shao No.2 as the female parent, Bozhou peony as the male parent to cross, collecting hybrid seeds, cleaning and disinfecting, taking embryos to obtain seed embryos; 2) taking the seed embryos, connecting into seed embryo starting seedling culture medium to culture, after the F1 generation seed embryo sprouts, first low-temperature treatment and then constant-temperature culture to obtain tissue culture seedlings; 3) taking the tissue culture seedlings to harden and transplant, which can be used. Under the culture of the specific culture medium, the double dormancy problem of the peony hybrid seeds is solved, and the low germination rate, high rot rate and embryo abortion of the peony hybrid seeds are overcome, so that the difficulty of the hybrid peony seedling quantity and quality is solved.

The invention discloses application of the ratio of phosphatidylserine to taurocholic acid. Research finds that compared with assisted reproduction crowds capable of obtaining available embryos after embryo culture, the concentration of phosphatidylserine in follicular fluid of assisted reproduction crowds without available embryos after embryo culture is remarkably reduced, which prompts that phosphatidylserine can be used as a marker for predicting developmental potential of assisted reproduction embryos; the prediction module is used for predicting whether available embryos can be obtained or not in advance in an egg taking stage of assisted reproduction; and the ROC curve verifies that the AUC of the PS (21: 220: 4) is 0.741, and the AUC of the ratio of the PS (21: 220: 4) to the taurocholic acid is 0.784, so that the accuracy and the sensitivity are higher when the ratio of the PS (21: 220: 4) to the taurocholic acid is used as the marker to predict whether available embryos exist or not.

The invention discloses an embryo development quality evaluation method and device, and belongs to the field of embryo in-vitro culture, the embryo before implantation is subjected to three-dimensional imaging to form a three-dimensional blastocyst structure chart, fluorescence imaging is carried out to form a redox specific heat chart, the three-dimensional blastocyst structure chart and the redox specific heat chart are subjected to co-localization superposition, and the embryo development quality is evaluated. Constructing a three-dimensional blastocyst model based on map combination; according to the three-dimensional blastocyst model, the spatial form, chromosome ploidy and metabolic state of the early blastocyst are comprehensively evaluated, and non-intrusive, accurate and efficient blastocyst ectodevelopment quality evaluation and transplantation screening are achieved.

The invention relates to a separation and purification method of yellow feather broiler chicken reovirus, a primer group for detecting virus and application. The ARV virus is detected from a sampled joint tissue of a sick chicken, the allantoic membrane has thickened leukoplakia and the characteristic cell fusion lesion can be seen through chick embryo culture, the purified virus can be observed through a transmission electron microscope to form circular virus particles with a double-layer membrane structure and the diameter of 60-80 nm, the circular virus particles accord with the morphological characteristics of the ARV, and the ARV virus can be used for preparing a medicine for treating the sick chicken. Animal regression experiments also prove that the separated ARV virus is a pathogen causing chicken arthritis. Genotype identification and whole genome sequencing are carried out after chick embryo rejuvenation and cell plaque purification, real-time fluorescent quantitative PCR universal primers and specific primers of all genotypes are designed according to a sequencing sequence, and efficient and accurate detection of the jute feather broiler ARV in the Xinjiang region is achieved.

This invention relates to a medaka hatching enzyme, its preparation method, and its application. The preparation method of the medaka hatching enzyme includes the following steps: 1) collecting fertilized medaka eggs; 2) incubating the fertilized medaka eggs with embryo culture medium in a 26℃ incubator for 6 days, and collecting mature embryos; 3) adding NaCl solution and Tris-HCl buffer (pH=8.0) to the mature embryos, grinding them, collecting the filtrate, and allowing it to stand overnight at 4℃; 4) centrifuging the filtrate at 15000 rpm for 15 minutes, and extracting the supernatant to obtain the medaka hatching enzyme solution. The preparation method is simple to operate, and the obtained medaka hatching enzyme has high activity, which can help to quickly remove the medaka eggshell artificially. The naked medaka embryos obtained by this method can be directly used for in vivo imaging and in-situ quantitative analysis, and embryo model construction, providing support for subsequent genetic, developmental biological, and toxicological studies of medaka embryos.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a biological response type memristor for in-vitro embryo monitoring and application of the biological response type memristor. The memristor comprises a substrate, a bottom electrode, a dielectric layer and a top electrode which are sequentially arranged from bottom to top, the dielectric layer is a double-layer functional layer composed of an Nb2O5 thin film and a barium titanate thin film, the barium titanate thin film is located on the upper layer of the bottom electrode, and the Nb2O5 thin film is located on the upper layer of the barium titanate thin film. The memristor realizes response to biological information by monitoring metabolic byproducts and oxidative stress markers in an embryo in-vitro culture solution in real time, realizes operation by inducing redistribution of surface charges of an interface through ion adsorption, and shows a resistance state transformation characteristic. Besides, the system breaks through the limitation of traditional microscope visual evaluation, realizes direct, sensitive and quantitative electric signal reading of the embryo metabolism state and the oxidative stress level, and solves the problem of embryo development potential evaluation caused by the increase of aneuploid rate in old lying-in women and recurrent implantation failure groups.

This invention provides an application and method of lysophosphatidylcholine (LPC) in promoting early development of reconstructed embryos, belonging to the field of embryo engineering technology. The invention first prepares reconstructed embryos, then adds lysophosphatidylcholine to the culture system for culturing. Lysophosphatidylcholine effectively promotes early development of the reconstructed embryos; at an optimal concentration of 5 μM, the blastocyst development rate increases to 24.56%, providing a novel and operable biochemical intervention target to solve the problem of low reconstructed embryo culture efficiency. This invention not only fills the gap in our understanding of the mechanism of action of LPC in this field, but also achieves controllability and reproducibility of the technical solution through precise definition of the optimal concentration. It provides key parameters for establishing standardized culture procedures and provides a more solid foundation for downstream applications such as developmental biology research and disease model construction that rely on high-quality reconstructed embryos, significantly accelerating the process of related scientific discovery and application development.

The invention belongs to the technical field of biology, and particularly relates to an embryo in-vitro rotating culture method. The method comprises the following steps: culturing an embryo in an embryo culture solution, wherein the embryo culture solution contains fresh rat serum, antibiotics, ascorbic acid or a derivative thereof, pyruvic acid or a salt thereof, glutamine, a buffer salt and optional glucose; the embryo culture solution does not contain human umbilical cord blood serum; the use of human umbilical cord blood serum is avoided, so that the embryo rotating culture system can be quickly popularized; the method has the advantages that the method is simple and easy to operate, and further limits and adopts fresh rat serum and adjusts the oxygen concentration in different periods, so that the embryo proportion of blood continuously flowing in the egg sac blood vessel in the E7.5 + 3day period can be increased, the rapid flowing of the blood in the egg sac blood vessel in the E7.5 + 5day period of the mouse embryo is realized, and the in-vitro culture process of the mouse embryo E7.5-E7.5 + 6day is completed.

A method for labeling embryo culture dishes used in assisted reproductive technology is provided, comprising the following steps: inserting the embryo culture dish into an adapter; inserting the adapter into the drawer of a labeling machine and closing the drawer; inputting or importing embryo-related information corresponding to the embryo culture dish into a host computer; and the labeling machine labeling the embryo-related information onto the embryo culture dish. This method allows for the automatic and rapid labeling of embryo-related information before use, facilitating accurate, efficient, centralized management and culture of large numbers of embryos for assisted reproductive technology.

The present invention discloses an ICSI operation method for improving the efficiency of producing sexed OPU embryos in vitro in dairy cows: frozen sexed sperm is treated using the BO flotation method, and the floated sperm is centrifuged and diluted with IVF. The treated sexed sperm is injected into OPU donor oocytes, activated twice with 5nM ionomycin and once with 2nM 6-DMAP, and then embryo culture is performed. The present invention significantly improves the blastocyst rate of embryos produced after ICSI eggs are activated, and can be used to stably produce high-quality sexed OPU embryos in dairy cows.

The embryo culture dish comprises a dish body and a dish cover, a plurality of groups of culture grooves are formed in the inner surface of the bottom of the dish body, the culture grooves in each group are not communicated, each group of culture grooves comprises a first hole groove and a second hole groove, the first hole grooves and the second hole grooves are in one-to-one correspondence and are communicated through through grooves, and the first hole grooves are communicated with the second hole grooves through the through grooves. Each first hole groove is connected with a second hole groove through a through groove, so that the situation that culture solutions of a plurality of embryos are mixed together to cause mutual pollution is avoided, and after culture is finished, the embryos in the first hole grooves are sucked away through suction pipes for detection or transplantation; the pipettor gun head is inserted into the bottom of the second hole groove, all the culture solution is slowly sucked out for noninvasive embryo detection, and the situation that due to inconvenient operation, mineral oil covering the upper portion of the culture solution easily pollutes the whole culture solution and interferes with follow-up detection or transplantation is avoided.