54 results about "Embryo freezing" patented technology

The present invention discloses embryo vitrifying freezing, defreezing and directly transplanting method, and belongs to the field of embryo biotechnology. The technological scheme includes setting freezing liquid and defreezing liquid in different sections of fine tube separated with air, setting embryo in one section of the freezing liquid, sealing ends of the fine tube, and throwing the fine tube into liquid nitrogen; and features that there are two sections of freezing liquid and two sections of defreezing liquid with volume ratio between the freezing liquid and defreezing liquid of 1 to 5, or antifreeze protecting agent concentration in the mixed liquid of 2.0 mol / L. The simple defreezing and transplanting process includes taking out the fine tube from liquid nitrogen, air bathing for 4-6 sec, water bathing at 20-25 deg.c, shaking for homogeneous mixing of freezing liquid and transplanting. The present invention realizes the simple tube detoxication of embryo and direct transplanting.

The invention discloses a lowline embryo freezing liquid, a freezing method and an unfreezing method and belongs to the technical field of bioengineering and embryonic heredity. The lowline embryo freezing liqui comprises ethanediol, cane sugar and DMSO. The freezing method comprises the following steps: freezing an obtained embryo for pre-treating, putting the embryo in a tube and freezing the embryo. The unfreezing method comprises the following steps: retaining a straw in air for 5-10 seconds, putting the straw in water bath at the temperature of 35-38 DEG C for 10-15 seconds, detecting the embryo, transferring the embryo in WM1 liquid drops, transferring into another hole liquid drops containing the same solution, then respectively transferring to WM2 and liquid drop containing HM with the concentration being 800 microliter per hole and retaining for 5 minutes each time; and washing the embryo by a culture solution for three times, continuously observing the development condition of the embryo in the culture solution or transplanting the embryo to a receptor cow after being subjected to estrus synchronization. The freezing liquid, the freezing method and the unfreezing method have the good freezing effect, are strong in operability, convenient to operate and suitable for popularization and application.

An enclosed type embryo freezing carrying rod comprises a carrying rod body, wherein an upper concave step and a lower concave step are arranged on the carrying rod body, a horizontally placed embryo slide is fixed on the lower concave step, a groove for holding embryos is arranged at the middle part of the embryo slide, and a cover plate connected to the carrying rod body and matching with the embryo slide is arranged on the upper concave step. The carrying rod body, the embryo slide and the cover plate form an enclosed type embryo carrier so as to avoid cross infection in a liquid nitrogen container; the carrying rod body is formed by injection molding, the embryo slide is formed by hot press molding, the production is convenient, and the fabrication cost is low; the embryo slide and the cover plate are both made of medical transparent PET thin sheet, and have good transparency and are easy for observation, and surface tension of the embryo slide after corona treatment is set to be 41-46 dynes, so as to avoid deformation of the embryo placed on the embryo slide; and after an embryo is placed on the embryo slide, the cover plate is hot sealed on the carrying rod body by pressing and hot sealing, the use is convenient.

The invention discloses an application of RhoA recombinant protein in preparation of a culture solution for improving embryo freezing resistance and recovery rate after thawing. The research finds that by addition of in vitro recombined RhoA protein to a culture solution of normally hatched embryos of mice, compared with a control group, the thawing recovery rate of normally hatched embryos can be significantly increased. The RhoA recombinant protein is applied to the field of embryo freezing for the first time, through addition of the RhoA recombinant protein, the freezing resistance and the recovery rate after thawing of the embryos are significantly increased, and use of a toxic cryoprotectant is reduced to a certain extent, so that the adverse effects on the embryos are reduced, and the RhoA recombinant protein has a wide application prospect in the technical field of cryopreservation of the embryos.

The invention relates to a refrigeration method for preserving sea fish embryo which comprises, selecting heartbeat stage embryos, charging into 0-4 deg C. antifreezing liquid, pre-cooling for 90-60 minutes, sectionwise cooling down, placing cores at -12 deg C during the cooling period, preserving in liquid nitrogen at the temperature of -30 deg C, equalizing 10 minutes at -80 deg C when defreezing, defreezing with 43 deg C water till full thawing, eluting with eluent, cultivating in sea water so as to obtain the revival embryos.

The invention belongs to the technical field of oocyte freezing, and particularly relates to a cryopreservation liquid and a cryopreservation method for oocytes. The cryopreservation liquid comprisesone part of an equilibrium liquid I, one part of an equilibrium liquid II and one part of one vitrification liquid. According to the invention, focused on the cell structure of oocytes, a specific freezing reagent for oocytes is innovatively designed by optimizing a conventional freezing system, improving the proportions of freezing components and adding a specific freezing protective agent; and through verification via mouse embryo and waste human oocyte freezing experiments, the specific freezing reagent for oocytes is proved to have greatly improved effects compared with existing embryo freezing reagents.

The invention relates to a drawer-type high-efficiency oocyte or embryo freezing loading rod, which includes a loading box, a carrier component and a marking plate. The marking plate is arranged on the loading box, and the loading box is side-opened Cubic structure, the interior of the loading box is divided into multiple freezing cavities by multiple partitions, the carrier assembly includes a connecting plate and a plurality of loading rods, one end of the loading rod is connected to the connecting plate, and the connecting plate of the carrier assembly is passed through a tight The fixing device is closely matched with the loading box body, and the loading rod is located in the freezing cavity. The invention has the advantages of improving storage space utilization and freezing efficiency of the freezing tank, and reducing the cost of purchasing liquid nitrogen tanks and liquid nitrogen; in addition, the loading rod is located in the freezing cavity and is completely protected by the loading box, and the embryo or oocyte Cells will not shake in liquid nitrogen, nor will it cause loss of embryos or oocytes; the marker plate has a larger area, which can mark more relevant information, improve the accuracy of recording, and is not easy to be confused.

The present invention provides a buffalo embryo freezing method in combination with laser membrane rupture. The method comprises: culturing an in vitro fertilized buffalo embryo to be an expanded blastocyst; using a laser membrane rupture instrument to drill a blastocyst cavity; discharging blastocyst fluid; and then using freezing liquid to perform vitrification freezing. According to the present invention, discharging the blastocyst fluid before freezing can make the cryoprotectant enter an embryo more quickly, reduce the possibility of ice crystal forming in a cell, and thereby avoiding embryo damage to a large extent, and improving a freezing survival rate of the embryo. On the other hand, due to partial breaking of a zona pellucida before freezing, so that after thawing cultivation, the zona pellucida is easier to be hatched in a revived blastocyst cavity, and thereby facilitating embryo implantation of the embryo after transplanting, and improving a transplant pregnancy rate.

The invention discloses a new application of gadol polysaccharide to prepare freezing deicing fluid of biological cell, mammal sperm, egg, protecting-body cell, early embryo and organ tissue, which comprises the following steps: allocating the basic liquid through glucose, Tris, sodium citrate and deionized water; making the rate of basic liquid d and yolk at 5-9:1-5; adding penicillin and streptomycin to obtain the liquid I; adding glycerin in the liquid I to produce liquid II; adding gadol polysaccharide in the liquid II to obtain the freezing deicing fluid of biological cell.

The present invention relates to a method for super-conventional breeding of fine-bred dairy cows using embryonic biotechnology, selecting excellent and suitable cows as donors, the method comprising the following steps: a, superovulation, b, artificial insemination c, embryo punching, d , Embryo inspection, e, embryo freezing, f, embryo thawing, g, embryo separation, h, embryo transfer. By adopting the above method, the birth rate of female calves can be effectively controlled, the production cost is reduced, the quality of dairy cows and milk is improved, and the purpose of supernormal rapid breeding of female dairy cows is achieved.

The invention discloses a wheat embryo freezing and storing method. According to the wheat embryo freezing and storing method provided by the invention, wheat embryos are stored at the temperature of minus 10 DEG C-0 DEG C. According to the wheat embryo freezing and storing method, immature wheat ears are stored at a low temperature below zero, which prolongs the supply time of the wheat embryos, and solves the problem to a certain extent that the achievement of the wheat embryo is limited by seasonality and artificial climatic conditions, so that the wheat embryo freezing and storing method has an important meaning to the large-scale trangenosis research of the wheat. Experiments show that minus 10 DEG C-0 DEG C is the temperature condition suitable for long-term wheat embryo storage, the wheat embryos can be stored for about 10 days at 10 DEG C, the regeneration rate of the wheat embryos has no significant difference from that of control group wheat embryos which are not stored at low temperature, and the real germination rate is obviously reduced; and the wheat embryos can be stored for about 25 days at -5 DEG C, the regeneration rate of the wheat embryos, especially the regeneration rate of the wheat embryos stored for 5-15 days, is higher than the regeneration rate of the control group wheat embryos which are not stored at low temperature, the generation rate of the wheat embryos stored at -5 DEG C for about 25 days is obviously different from the regeneration rate of the control group wheat embryos, and meanwhile the real germination rate is obviously lowered.

The invention belongs to the field of embryo cryopreservation and particularly relates to an in-vitro embryo culture solution for increasing the embryo unfreezing thawing rate. The in-vitro embryo culture solution comprises Clusterin protein. The invention further relates to a method for improving embryo freezing resistance and increasing the unfreezing thawing rate and a method for freezing an embryo. By means of the in-vitro embryo culture solution and the methods, the freezing resistance of the embryo can be remarkably improved, and the unfreezing thawing rate of the embryo can be remarkably increased.

The invention relates to a telescopic efficient oocyte / embryo freezing carrying rod. The telescopic efficient oocyte / embryo freezing carrying rod comprises a carrying rod part and a carrying rod sleeve pipe; the carrying rod part consists of a handheld rod and a freezing blade; the handheld rod is a telescopic rod; the telescopic rod consists of 2 to 8 hollow pipes with different diameter in a sleeving way; and one end of the telescopic rod is fixedly connected with the freezing blade. A cold tool has the following advantages: 1, the handheld part of the carrying rod adopts a telescopic rod structure, the step of putting the freezing carrying rod into a freezing bracket by hands can be guaranteed, the handheld part is long enough and convenient to operate, the handheld part is shortened toone third or one fourth of the original freezing carrying rod after the freezing carrying rod is put into the freezing bracket, one bracket can store the freezing carrying rods with the number whichis three times or more times that of the original number, and the utilization efficiency of a freezing tank can be increased by about three times; and 2, the expenditure of buying a liquid nitrogen tank and liquid nitrogen can be greatly saved and the freezing cost of the oocyte / embryo is reduced.

The invention discloses a vitrification freezing solution for an embryo. According to the vitrification freezing solution for the embryo, the three agents of hydroxypropyl cellulose (HPC), human serumalbumin (HSA) and glycerinum are added to the freezing solution to have a combined effect, and the better capacity of resisting damage is achieved. After the embryo is treated, frozen and stored in the vitrification freezing solution, the blastulation rate of the in-vitro embryo after the embryo is unfrozen can be increased. According to the vitrification freezing solution, the blastulation rateafter the embryo is unfrozen can be greatly increased.

The invention discloses an unfreezing liquid for frozen equine animal embryos, and a preparation method and application thereof. The unfreezing liquid includes a sodium lactate mixed solution, penicillin, streptomycin, and one, two or three selected from bovine serum albumin, calf serum and inactivated equine serum. The preparation method includes the steps of: placing the sodium lactate mixed solution in a thermostat at 37 DEG C for 0.5-6 h for standby; and adding penicillin, streptomycin and one, two or three selected from bovine serum albumin, calf serum and inactivated equine serum into the sodium lactate mixed solution in accordance with the component contents, so as to obtain the unfreezing liquid for frozen embryos. The unfreezing liquid is suitable for non-surgical method for embryo transplantation of horses, cattle and donkeys. The invention has the beneficial effects of convenience for material acquisition, low price, simple preparation and convenience for usage, is very suitable for scale production of non-surgical method for embryo transplantation of equine animals, and is favorable for popularization and application of embryo transplantation technology.

The invention relates to a method for unconventionally breeding improved cows through an embryo biotechnology. Excellent and suitable female cows are selected as donors. The method comprises the following steps of (1) superovulation, (2) artificial insemination, (3) embryo washing, (4) embryo checking, (5) embryo freezing, (6) embryo unfreezing, (7) embryo separation, and (8) embryo transplantation. By the adoption of the method, the birth rate of female calves can be effectively controlled, production cost is reduced, quality of the cows and milk is improved, and the purpose of unconventionally and rapidly breeding the cows is achieved.

The present invention discloses embryo vitrifying freezing, defreezing and directly transplanting method, and belongs to the field of embryo biotechnology. The technological scheme includes setting freezing liquid and defreezing liquid in different sections of fine tube separated with air, setting embryo in one section of the freezing liquid, sealing ends of the fine tube, and throwing the fine tube into liquid nitrogen; and features that there are two sections of freezing liquid and two sections of defreezing liquid with volume ratio between the freezing liquid and defreezing liquid of 1 to 5, or antifreeze protecting agent concentration in the mixed liquid of 2.0 mol / L. The simple defreezing and transplanting process includes taking out the fine tube from liquid nitrogen, air bathing for 4-6 sec, water bathing at 20-25 deg.c, shaking for homogeneous mixing of freezing liquid and transplanting. The present invention realizes the simple tube detoxication of embryo and direct transplanting.

The invention belongs to the technical field of chemicals delivery. In order to solve the problems of difficulty in delivery operation and poor stability of chemicals when the chemicals are delivered during ovum / embryo freezing treatment in the prior art, the invention discloses a chemicals delivery system and a method. The system comprises a carrier and a chip. A groove is formed in the carrier and is used for containing solution. An interval is arranged on the chip, and internally provided with chemicals to be delivered. The coverage area of the interval is larger than the opening area of the groove. The chip and the carrier can move relative to each other. The chemicals to be delivered located in the interval can make full-coverage contact with the solution in the groove, and forms free diffusion between the chip and the carrier. According to the present invention, when the chemicals are delivered during ovum / embryo freezing treatment, the delivery operation difficulty can be lowered, and the stability and reliability of the operation process are improved.

The invention provides a bovine cloned embryo freezing solution, a bovine cloned embryo thawing solution, a kit and bovine cloned embryo freezing and thawing methods, and belongs to the in-vitro embryo freezing technology. The freezing method comprises the following steps: putting a bovine cloned embryo into a basic solution, and balancing the embryo at a temperature of 24-25 DEG C for 5-10 minutes; putting the processed embryo into a first vitrification freezing solution to balance the embryo at a temperature of 24-25 DEG C for 4-6 minutes, putting the processed embryo into a second vitrification freezing solution to balance the embryo at a temperature of 24-25 DEG C for 4-6 minutes and putting the processed embryo into a third vitrification freezing solution to balance the embryo at a temperature of 24-25 DEG C for 35-45 seconds, and finally putting the processed embryo into liquid nitrogen to freeze. And the frozen bovine cloned embryo is put in a first thawing solution, the embryois thawed at a temperature of 37-39 DEG C for 50-70s, the thawed embryo is put in a second thawing solution, the embryo is balanced at a temperature of 24-25 DEG C for 4-6 min, and finally placing theprocessed embryo is put in a base solution, and the embryo is balanced at a temperature of 24-25 DEG C for 4-6 min. With the method disclosed by the invention, the conception rate of the bovine cloned embryo is as high as 25%.

The invention discloses a freezing and unfreezing control system for human embryo cryopreservation. The system comprises a box body, a computer main control system, a display system, a pipeline systemand a micro camera system are arranged in the box body, the computer main control system controls the whole process of embryo freezing and unfreezing, the pipeline system comprises a liquid nitrogenconveying pipe and a plurality of pipelines, one end of the liquid nitrogen conveying pipe is connected with a liquid nitrogen controller, one end of each one of the pipelines are connected with a refrigerating fluid storage bottle, an unfreezing fluid storage bottle and a waste liquid storage bottle, and the liquid nitrogen conveying pipe and the other ends of the pipelines are connected with a first needle type probe, a plurality of drawer type units are arranged on the inner wall of the box body, a micro cryopreservation device is arranged in the drawer type unit, a movable track is arranged in the box body, the micro camera system is arranged on the track and moves and rotates along the track at regular time so as to shoot embryo images in a micro cryopreservation device, and the shotembryo images are displayed through a display system. Integrated operation can be carried out, repeated transfer is not needed, and multiple embryos can be frozen at the same time.

The invention belongs to the technical field of chemical delivery. In order to solve the problems that operation is inconvenient and an ovum / embryo is flushed out of a carrier or directly sucked out when an ovum / embryo and a solution are subjected to contact treatment by adopting a suction tube, the invention discloses a method for feeding a solution to a biological material by utilizing hydrogel. The method specifically comprises the following steps: S1, preparing a solution to be put into a hydrogel structural form; S2, contacting a biological material with the hydrogel prepared in the step S1, and completing the feeding operation through diffusion of a solution in the hydrogel to the biological material. When the method provided by the invention is adopted to carry out the solution feeding operation in the ovum / embryo freezing treatment process, not only can the risk of flushing out or sucking out the embryo possibly caused when a suction tube is used for feeding and sucking out the solution be completely avoided, but also the reliability and the convenience of the feeding operation can be improved, and the normal proceeding of subsequent freezing treatment is ensured.

The invention discloses an auxiliary safe docking device of human embryo freezing carrier poles under liquid nitrogen and a using method of the auxiliary safe docking device. According to the auxiliary safe docking device of the human embryo freezing carrier poles under the liquid nitrogen and the using method of the auxiliary safe docking device, disclosed by the invention, limiting and fixing are carried out by nesting protection caps of freezing carrier poles in cylindrical holes, meanwhile, the freezing carrier poles are arranged in cross grooves and are in face-to-face docking, the wholedocking process can be completed in the cross grooves formed in preset paths, docking observation under the liquid nitrogen requires not to be carried out by naked eyes, one-time success can be ensured, potential damage of a traditional method to embryos or staff can be effectively avoided, the operation is simple and convenient, and an effect is reliable.

The invention relates to a preservation method for human body, animal or its part and its freezing carrier, in particular to a vitrification freezing preservation method of oocyte / embryo and its freezing carrier. The method comprises: a freezing liquid preparation step, a thawing liquid preparation step, a freezing step and a thawing step, wherein the freezing carrier used in the freezing step and the thawing step is a freezing net, and the freezing net comprises: a mesh body and net handle. The invention provides a simple, fast and efficient oocyte or embryo vitrification freezing method and its freezing carrier.

The embryo freezing fresh-keeping device comprises a fresh-keeping tank, a first motor is arranged in the center of the upper portion of the fresh-keeping tank, the first motor is connected with a rotating frame, a limiting rod is vertically installed on the rotating frame, the limiting rod is sleeved with a test tube containing cylinder, and an automatic lifting device is arranged on one side of the upper portion of the fresh-keeping tank. A second motor is arranged in the automatic lifting device, a lifting lead screw penetrating through the guide rail groove is connected to the lower portion of the second motor, the lifting lead screw is connected with a grabbing mechanism in a matched mode through a lead screw nut sliding block, a taking-out opening is formed in the portion, below the grabbing mechanism, of the fresh-keeping tank, a threaded sealing cover is installed on the taking-out opening, and an opening-closing rotating disc is welded to the threaded sealing cover. According to the automatic lifting device for the fresh-keeping tank, through the arrangement of the automatic lifting device, the test tube placement barrel in the fresh-keeping tank can be automatically lifted out, manual lifting is omitted, the chance that a user makes contact with liquid nitrogen is reduced, and safety is greatly improved.

The invention discloses a storage device for accurately selecting frozen embryos for embryo freezing technology, which comprises a freezing tank, a cover plate rotatably arranged on the freezing tank, a handle for rotating the cover plate, and a handle rotatably arranged on the freezing tank. The sleeve on the cover, the pull rod rotatably arranged on the sleeve, the first support rod symmetrically arranged on the cover, and the freezing bracket arranged on the first support rod; The freezing support includes a support ring, a second support rod symmetrically arranged on the support ring, fastening grooves respectively arranged on the second support rods, and parallel grooves arranged between the first V-shaped guide rods , the second freezing tube arranged on the parallel groove. In the present invention, by adding a pull rod to the cryogenic tube, the required frozen embryos can be precisely selected without opening the frozen tube, so that other embryos will not be exposed to normal temperature and the quality of the embryos can be guaranteed.

The invention belongs to the technical field of embryology. The invention provides beef cattle reproductive embryo cryopreservation liquid and a freezing method. The cryopreservation liquid is prepared from the following components in parts by weight: 40 to 50 parts of an HPES buffer solution, 0.1 to 0.5 part of bovine serum albumin, 2 to 5 parts of sorbitol, 0.01 to 0.02 part of glutamine, 10 to15 parts of polyethyleneimine and 20 to 25 parts of cane sugar. The beef cattle reproductive embryo freezing method comprises the following steps: balancing beef cattle embryos to be frozen in a pre-balancing solution for 5-8 minutes, balancing the beef cattle embryos in the beef cattle reproductive embryo freezing preservation solution according to any one of claims 1-3 for 30-40 seconds, transferring the beef cattle reproductive embryo freezing preservation solution to a freezing carrier, and putting the freezing carrier into liquid nitrogen for freezing treatment. By means of the technicalscheme, the problem that in the prior art, the cryopreservation effect of cryopreservation liquid on cells is poor, and consequently the cryopreservation survival rate of the cells is low is solved.

The invention discloses an adjustable embryo freezing tubule scaleplate rack. The adjustable embryo freezing tubule scaleplate rack comprises an open type frame defined by two side walls, a bottom plate and a top plate, wherein an opening and closing controllable lock catch is arranged between the top plate and one side plate; an upper arc press sheet and a lower arc press sheet which are matched with the outer diameter of an embryo tubule are arranged in centers of the top plate and the bottom plate respectively; and the upper arc press sheet and / or the lower arc press sheet are / is provided with scaleplates. According to the invention, a frame structure for covering and fixing the embryo tubule is designed, and the corresponding scaleplates are arranged and can be combined with the embryo tubule well; not only can an imbibition reference standard be provided for the embryo tubule conveniently, but also the support strength of the embryo tubule can be improved, and shaking is prevented; the outer frame is light in weight, has certain hardness, can adopt transparent or hollow-out structure, and facilitates observation; and the adjustable embryo freezing tubule scaleplate rack is reasonable in structural design, good in use effect and suitable for popularization and application.

The invention discloses an integrated embryo freezing device which comprises an embryo freezing box, a plurality of loading plates and a cover plate. A plurality of partition plates are uniformly fixed in the embryo freezing box from left to right, so that the embryo freezing box is divided into a plurality of freezing chambers; the loading plates are uniformly fixed to the back end surface of thecover plate and are opposite to the freezing chambers in position, and embryo loading grooves are formed in the upper end surfaces of the loading plates, separately. The integrated embryo freezing device has the beneficial effects that the loading plates for loading embryos are integrated to the cover plate which is taken as a supporting plate shared by the loading plate without a handle, so thatthe occupied space can be reduced compared with scattered arrangement of a plurality of conventional carrier rods, the liquid nitrogen tank storage efficiency can be improved greatly and the liquid nitrogen tank storage space is fully utilized when the cover plate is placed on a freezing rack in the liquid nitrogen storage tank. The embryo loading grooves can load embryos needing to be frozen conveniently, so that the embryos are hard to slide and fall off. Therefore, the integrated embryo freezing device is good in using effect.

The invention discloses a cryopreservation method for embryos of Litopenaeus vannamei, which includes the steps of freezing embryos and thawing embryos; in the freezing of embryos, the embryos of Litopenaeus vannamei hatched for 8 hours are antifreeze firstly. Then the embryos are frozen by slow freezing method or quick freezing method; the thawing of the embryos is to take out the thin tube from the liquid nitrogen, quickly immerse it in water at 30°C, and elute it immediately after the thin tube becomes transparent. Embryos are rinsed into Petri dishes and placed in incubation buckets to incubate. According to the characteristics of Litopenaeus vannamei, this method prepares suitable antifreeze solution and eluent, and optimizes the process of antifreeze treatment and thawing treatment. The long-term needs of shrimp farming and research, and simple operation, easy to popularize and apply.

The invention relates to a refrigeration method for preserving sea fish embryo which comprises, selecting heartbeat stage embryos, charging into 0-4 deg C. antifreezing liquid, pre-cooling for 90-60 minutes, sectionwise cooling down, placing cores at -12 deg C during the cooling period, preserving in liquid nitrogen at the temperature of -30 deg C, equalizing 10 minutes at -80 deg C when defreezing, defreezing with 43 deg C water till full thawing, eluting with eluent, cultivating in sea water so as to obtain the revival embryos.