54 results about "Embryo freezing" patented technology

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.

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 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 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 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.