51 results about "Acrosome" patented technology

The invention relates to a diluent of frozen semen of a milk goat, and a preparation method and a diluting method thereof as well as a preparation method of frozen semen of the milk goat in a thin pipe. In an existing frozen semen storage method, sperms are struck by low temperature to cause structure change of plasmalemma, so that the integrity of the plasmalemma and the survival rate and the fertility of the sperms after resuscitation are seriously influenced. The preparation method is characterized in that thin-pipe freezing treatment with two-step dilution and balancing and two-step cooling is carried out on fresh semen, wherein a basic diluent is prepared from Tris, citric acid, glucose, trehalose, Vc, GSH, BSA, ATP, compound antibiotic, yolk and water; a frozen diluent is prepared from the basic diluent and glycerinum; the diluent is utilized for diluting and balancing the fresh semen, and the fresh semen is sub-packaged in thin pipes and is stored in liquid nitrogen. According to the invention, the effects of low-temperature storage and freezing storage of the frozen semen of the milk goat is good, the survival rate, the integration rate of acrosome and the integrity of the plasmalemma of the semen after unfreezing are effectively increased, the aberration rate is reduced and the production efficiency and the quality of the frozen semen of the milk goat are improved.

The invention discloses a novel frozen semen diluent used for improving the freeze-thaw quality and fertilization rate of frozen bovine semen and a preparation method thereof. Every 100ml of the prepared frozen semen diluent contains 0.9-1.2g of sodium citrate, 3-5g of sucrose, 18-22ml of yelk, 4-7ml of glycerin, 6-9mg of vitamin E, 500-700mg of vitamin C, 35mg of spectinomycin and 80 thousand units of gentamicin. The invention adds VE and VC into the semen diluent for the first time, so as to increase the motility rate and acrosome integrity of the frozen sperm as well as the activity of antioxidant enzyme in seminal plasma, thus improving the quality and fertilization rate of the freeze-thaw sperm.

The invention discloses an anti-freezing agent and an anti-freezing diluent for freezing and storing livestock sperm and a preparation method thereof. The anti-freezing agent for freezing and storing bovine and porcine sperm is algin, the amount of the algin added into the conventional diluent is 60 to 110 milligrams, wherein the conventional diluent is prepared from 1.1 grams of glucose, 1.48 grams of citric acid, 2.42 grams of Tris, 10 percent of fresh egg yolk, 0.06 gram of penicillin sodium and 0.1 gram of streptomycin sulfate, which are dissolved in 100 millimeters of distilled water. The adding amount of the algin for cattle is 100 to 110 milligrams, and the adding amount of the algin for pigs is 60 to 80 milligrams. The preparation method comprises the following steps of: adding 100 to 110 milligrams of algin and 4 percent glycerin (for cattle) or 60 to 80 milligrams of algin and 1 percent glycerin (for pigs) into the conventional diluent to prepare the anti-freezing diluent; and adjusting the pH value of the solution to be between 6.5 and 7.5, filtering and sterilizing the anti-freezing diluent under the osmotic pressure of 286mOsm / L, and putting the anti-freezing diluent into a 4 DEG C refrigerator for later use. The algin is used for freezing and storage of the bovine and porcine sperm for the first time, and after unfreezing, the sperm motility rate, the acrosome completeness rate and the plasma membrane completeness rate are 65 percent, 69 percent and 72 percent (for cattle) respectively and 58 percent, 62 percent and 63 percent (for pigs) respectively.

The invention discloses an albumen microsphere conjugate for detecting acrosin activity and a preparation method and application of the albumen microsphere conjugate. The albumen microsphere conjugate is characterized in that monodisperse micron microspheres obtained by combining fluorescein and the acrosome reaction substrate, namely zona pellucida protein are prepared, the acrosin activity is detected by aid of change in fluorescence intensity, and the specific steps include preparation of monodisperse microspheres modified through surface carboxylation or amination, surface modificaction, fluorescence labeling of protein polypeptide, conjugation of fluorescent protein polypeptide and functional microspheres and detection of the acrosin activity. The albumen microsphere conjugate has the advantages that protease digestion on the microspheres is realized through the polymer microsphere technology and the protein labeling and conjugating technology, and enzymatic activity is evaluated; the albumen microsphere conjugate is mainly applied to flow detection of the acrosin activity, synchronous detection of an acrosome reaction and the acrosin activity can be realized, and acrosome functions can be evaluated more truthfully and accurately.

The invention relates to a sperm acrosome zone Raman spectrum peak and a use thereof. The Raman spectrum peak is represented by an abstract figure shown in the specification, and can be used for the fertilization function assessment and detection and the theory researches of a sperm. The invention also relates to a sperm fertilization function assessment method. The method comprises the following steps: 1, preparing a spermatic slice; 2, scanning the single sperm acrosome zone on the spermatic slice at a laser excitation wave of 532nm under a laser power of 5mW three times to obtain a Raman data spectrum peak; and 3, contrasting the Raman data spectrum peak of a detection sample with the Raman data spectrum peak of the above sperm acrosome zone to determine that whether the sperm has a fertilization function or not. The method provides a standard Raman spectrum peak for determining the sperm fertilization potential, and the determination result of the method is accurate and reliable, and has a high credibility, so the method is a new sperm fertilization potential detection method; and a separate sperm can be controlled by the method, so there is no need to specially process the sperm, the damage of the sperm can controlled in an extremely low range, and the sperm can be directly used for subsequent researches or auxiliary reproduction treatment.

The invention relates to a noninvasive sperm form and ultrastructure analysis method. The method comprises the following steps: analyzing by using a dispersion type confocal Raman spectrometer, selecting an excitation Raman spectrum, placing sperm cells under a reflected light microscopy, and acquiring the chemical group spectrum of the sperm cells through a certain excitation power and a certain exposure time; respectively dividing the acrosome, the neck and the tail of a sperm into scanning areas, scanning three times to obtain a Raman data spectrum peak, and analyzing the waveform change and the displacement of the spectrum peak; and retrieving the acquired sperm form Raman spectrums in an RRUFF Raman database to obtain the Raman spectra of all areas of the sperm form. The laser Raman spectrum adopted in the invention is a noninvasive technology, so the characteristics of the ultrastructures of different individuals and the scattering substances of different components of each sperm can be reflected without any dye or fixation, and a new assessment method is provided for the sperm morphology analysis.

The invention relates to a non-animal-origin frozen semen diluent, and relates to a frozen semen diluent prepared with non animal-origin component. The diluent is used for preparing cattle and sheep frozen semen. In prior arts, adopted frozen semen contains animal-origin components, and prepared frozen semen has the defects of low motility and short survival time, low intact acrosome rate of thin tube frozen semen, and large amount of pathogenic micro-organisms after thawing. With the diluent provided by the invention, the problems are solved. The diluent is prepared from the components of, by mass, 3.412 g of lactose, 0.5 g of sucrose, 0.9 g of glucose, 0.61 g of trehalose, 0.53 g of citric acid, 0.403 g of acetic acid tetraethylammonium, 1.425 g of tris(hydroxymethyl) aminomethanegrams, 0.375 g of tri-sodium citrate, 5.2 ml of ethylene glycol, 100 ml of double-distilled water, and 0.88 g of phospholipids powder. With the diluent, prepared frozen semen has the advantages of high motility and long survival time, high intact acrosome rate of thin tube frozen semen, and small amount of pathogenic micro-organisms after thawing.

The invention discloses a sperm quality evaluation method based on flow cytometry, which comprises the following steps of: firstly, commonly dyeing a semipermeable membrane nucleic acid fluorescent dye, a non-semipermeable membrane fluorescent nucleotide dye and a sample to be detected; and then, incubating a fluorescent marker monoclonal antibody and the dyed sample to be detected and carrying out flow cytometric detection to obtain the final detection results,. The fluorescent marker monoclonal antibody can be reacted with a sperm acrosome membrane. The laser excitation wavelength differences between the fluorescent marker monoclonal antibody and the semipermeable membrane fluorescent nucleotide dye and between the fluorescent marker monoclonal antibody and the non-semipermeable membrane fluorescent nucleotide dye are 100-200 nm, and the fluorescent scattering wavelength differences are 75-150 nm. The sperm quality evaluation method has the beneficial effects through one-time detection, a plurality of parameter results can be simultaneously obtained, so that the apoptosis rate, the plasma membrane integrity rate and the survival rate of sperm are obtained, and the detection labor strength is reduced; and the demands on a large batch of sample detection in the clinical can be satisfied.

The present invention discloses applications of the compatibility of lycopene and sesamol in livestock semen freezing storage agents, and provides a livestock semen freezing storage agent, which can significantly improve the survival rate of livestock sperms, and comprises a freezing base liquid, egg yolk, lycopene and sesamol. The present invention further provides another livestock semen freezing storage agent, which comprises a freezing base liquid, egg yolk, lycopene, sesamol and glycerol. According to the present invention, the compatibility of lycopene and sesamol is firstly used as the anti-freezing agent for the freezing storage of the semen of pigs, sheep and cows, and the obtained livestock semen freezing storage agent can be used for the artificial insemination of the test tube freezing semen of pigs, sheep and cows, and the long-term storage and the after-long-distance-transportation semen deposition of the semen after the super-temperature freezing, and can significantly improve the survival rate, the acrosome integrity and the plasma membrane integrity of the frozen-thawed perms of pigs, sheep and cows,.

The invention discloses a pig feed capable of improving the quality of breeding boar semen fluid and a preparation method thereof. The feed is prepared from the following materials by weight parts: 550-600 parts of corn, 100-150 parts of puffing bean pulp, 200-250 parts of bran, 40-50 parts of a premix, 10-15 parts of calcium hydrophosphate, 3-5 parts of salt, 2-3 parts of sodium bicarbonate, 3-5 parts of attapulgite, 5-8 parts of silene aprica, 4-6 parts of herba epimedii, 2-3 parts of pumpkin seeds, 3-4 parts of viscum album, 1-3 parts of tuber onion seeds, 1-2 parts of human placenta, 2-3 parts of semen cuscutae, 10-15 parts of chestnut shells and 5-10 parts of spartina anglica. The feed prepared by using the method disclosed by the invention not only can provide various nutrients needed by the healthy growth of breeding boars and has good feed palatability, but also can obviously improve the semen volume, the sperm density and the sperm survival rate; meanwhile, the acrosome abnormal rate and the rate of sperm deformation are obviously reduced.

The invention discloses an acrosome reaction kit, which comprises tissue slide preserving fluid, a fluorescent conjugate and a fluorescent sealing solid liquid, wherein each 100 ml of the tissue slide preserving fluid contains 0.5ml of glutaraldehyde, 5ml of methanol, 2ml of ethanol, 0.2g of polyvinylpyrrolidone (PVP), 1ml of formalin and the balance of purified water. The kit can carry out a sperm-induced acrosome reaction and detection; and if calcium ion induction is not carried out, the kit can carry out the acrosome reaction condition evaluation and the acrosome integrity determination. By the tissue slide preserving fluid, the test process for the sperm-induced acrosome reaction and detection is carried out by two parts, samples and test results which are obtained in the first part can be preserved and the final test result cannot be influenced if the operations of the second part are carried out in the subsequent 72 hours, so that the work intensity can be relieved. The fluorescence of a fluorescent staining system is strong and cannot be attenuated after being stabilized for 15 days, so that the detection result is more stable and is easy to interprete and the repeatability of the result can be ensured. The kit and a determination method can be clinically applied and popularized.

The invention provides a livestock seminal fluid cryoprotectant and an application thereof. The cryoprotectant comprises a fluid A and a fluid B, wherein the fluid A includes glucose, edetic acid, dihydrate sodium citrate, sodium bicarbonate, potassium chloride, penicillin and streptomycin, and the fluid B includes fructose, tris hydroxymethyl aminomethane, citric acid, penicillin, streptomycin, glycerol, DMF (Dimethyl Formamide), glycol and yolk. According to the livestock seminal fluid cryoprotectant, frequently-used three refrigerants of glycerol, glycol and DMF are optimized and combined, so that an optimum compatibility effect of the three cryoprotectants is achieved. The selected cryoprotectant composition capable of improving a cryopreservation effect of the livestock seminal fluid can effectively improve the cryopreservation survival efficiency of the livestock seminal fluid, can maintain the integrity of the acrosome, can be widely popularized and applied to the field of livestock cultivation, and has favorable market effects and social effects.

The invention discloses an application of a polysaccharide substance in preparation of a dilution solution for freeze preservation of goat sperms and a preparation method of the dilution solution. Thepolysaccharide substance comprises one or more selected from the group consisting of tremella polysaccharide, lentinan, auricularia polysaccharide and cordyceps polysaccharide. The invention also provides the dilution solution for freeze preservation of the goat sperms. The dilution solution comprises the polysaccharide substance, a nutrient substance, a buffer substance and the like. The dilution solution for freeze preservation of the goat sperms provided by the invention has the effects of improving a sperm motility rate, an acrosome and plasma membrane integrity rate after freeze preservation, and significantly improving goat reproductive performance and economic benefits.

The invention discloses an application of phloretin in preparation of a pig, sheep or bovine semen cryopreservation agent. Meanwhile, the invention discloses a cryopreservation agent containing phloretin and a freezing base solution. According to the invention, phloretin is used for cryopreservation of semen of pigs, sheep and cattle for the first time; the application is reliable in effect, easyto popularize and suitable for artificial insemination of pig, sheep and cattle tubule frozen semen, long-term storage of the semen after ultralow-temperature freezing and semen deposition after long-distance transportation, the sperm motility rate, acrosome integrity rate and plasma membrane integrity rate of the frozen-thawed pig, sheep and cattle can be remarkably increased, and the fertilization capacity of sperms is kept.

The invention discloses a normal-temperature preservation antioxidant diluent for swine semen and a preparation method thereof. The diluent is prepared from glucose, EDTA, potassium chloride, disodiumcitrate, sodium bicarbonate, gentamicin and isatis root polysaccharide. The plant-derived natural antioxidant isatis root polysaccharide is used for replacing an artificially synthesized antioxidant,so that the antioxidant capacity of swine semen can be enhanced, oxidative damage can be relieved, the motility rate, plasma membrane integrity and acrosome integrity of the swine semen can be effectively maintained, and finally, the fertilization capacity of the semen can be maintained. The normal-temperature liquid storage time of the swine semen can be prolonged, and the requirements of pig production in remote areas are satisfied; meanwhile, the diluent is low in cost, simple to operate, stable in preservation effect and suitable for wide popularization. The technology has important significance for promoting healthy and sustainable development of the pig industry.

The invention discloses a protective agent applied to frozen storage of livestock semen. The protective agent can remarkably improve the survival rate of livestock sperms, and the main substance of the protective agent is ganoderma lucidum polysaccharide. The preparation method is mainly characterized in that the ganoderma lucidum polysaccharide is used as the protective agent for long-term preservation of straw frozen semen of pigs, sheep and cattle, for artificial insemination, and for insemination after long-distance transportation of the semen subjected to ultralow temperature refrigeration for the first time at home and abroad. The survival rate, the acrosome integrity and plasma membrane integrity of sperms of pigs, sheep and cattle after freezing and thawing are notably improved.

The invention relates to a freezing preservation method of boar semen, which comprises the following steps: firstly preparing a freezing dilution liquid I, a freezing dilution liquid II and a dilution liquid BTS, then carrying out hydrotreating and semen collection on the three solutions, diluting the boar semen with the hydrotreated dilution liquid BTS at the constant temperature of 1: 1, balancing at 17 DEG C for 2-4 hours, and centrifuging to remove the supernatant, thereby obtaining the boar semen. The method comprises the following steps: diluting sperm precipitates by using an isothermal hydrotreated frozen dilution liquid I, then cooling and balancing to 4 DEG C, finally diluting by using an isothermal hydrotreated frozen dilution liquid II according to a ratio of 1: 1, sub-packaging the diluted sperm by using frozen tubules, sealing, freezing in a box body of a programmed freezing instrument, and after freezing is finished, storing the frozen sperm in a refrigerator. And transferring the frozen tubule into liquid nitrogen for storage. The method is simple to operate, can effectively improve the unfreezing activity and the sperm acrosome intact rate of the frozen pig sperms, can reduce the oxidative damage degree of the sperms, and overcomes the problem that the sperms are easily subjected to oxidative damage in a long cooling balance process.

The invention provides a formula and a dilution method of a diluent for normal-temperature preservation of Hu sheep semen. According to the invention, Hu sheep semen is subjected to normal-temperaturepreservation by using five diluents, and the motility rate, the vitality, the plasma membrane integrity rate, the acrosome integrity rate, the survival time and a certain motion parameter in the semen preservation period are detected by using a computer-aided sperm analyzer (CASA), a Coomassie brilliant blue staining method, a hypotonic test method and other methods; according to the diluent formula obtained by screening, the motility rate and the vitality are slowly and stably reduced in the semen preservation period, the time at the vitality of more than 0.6 can reach more than 3 days, andthe total survival time can reach more than 17 days; and a purpose of the invention is to objectively evaluate and screen out a formula of a diluent with the best Hu sheep semen normal-temperature preservation effect so as to provide reference for popularization and application of a Hu sheep semen preservation technology in production and further improve the breeding scale of Hu sheep.

The invention discloses a novel cryoprotective liquid, which comprises: 200mL / L of yolk, 13.536g / L of sodium citrate, 24.224g / L of trihydroxymethyl aminomethane, 11g / L of glucose, 1g / L of 1 million unit streptomycin sulfate, 0.8g / L of 1.6 million unit penicillin, 30mL / L of glycerin and 0.05moL / L-0.15moL / L of trehalose. According to the cryoprotective liquid, the toxicity of the protectant is reduced, and the freeze-thawing effect is improved; when sperms are frozen, the vitality, plasma membrane integrity, acrosome integrity, tyrosine phosphorylation level, protamine deficiency level, motilityrate and mitochondrial membrane potential of the sperms can be remarkably improved, and the quality of the frozen and thawed sperms is improved.

The invention provides application of 2-o-mercapto-acetic indican ascorbic acid in preparation of semen for controlling gender, which has the following advantages that AA-2G is applied to a preparation process of the semen for controlling gender to inhibit a large amount of free oxygen generated in the preparation process; and because the AA-2G is stable in water solution, the damage to the semen caused by treatment processes of separation, centrifugation and the like in the preparation process can be avoided to the maximum, thereby obviously improving semen quality parameters, such as semen activity, acrosome completeness and plasmalemma completeness, and the 2-o-mercapto-acetic indican ascorbic acid is obviously prior to the traditional anti-oxidizing agent with respect to action effect and action time.

The invention discloses an application of heparin sodium in preparation of a diluent for goat sperm cryopreservation and a preparation method, and also provides a diluent for goat sperm cryopreservation, which comprises heparin sodium, nutrient substances, buffer substances, a cryoprotectant and antibacterial substances. The diluent for goat sperm cryopreservation plays a role in protecting spermsin the processes of sperm cooling, cryopreservation and unfreezing, damage is reduced, and the sperm motility and acrosome integrity rate of goat frozen sperms are obviously improved.

Provided is a preparation method and application thereof for dog seminal fluid cryopreservation diluent. The diluent comprises diluent A, diluent B, diluent C and diluent D, wherein the diluent A comprises vitamin E, glucose, citric acid, fructose, yolk liquid, carnitine, sodium glutamate, penicillin, and streptomycin; the diluent B comprises glucose, DMSO, glycerinum and glycol; the diluent C comprises mycose, gelatin, EDTA, sorbitol, polyvinyl pyrrolidone and peptone; the diluent D comprises thiomersalate and caspase inhibitor Z-VAD-FMK. According to the prepared diluent, the sperm motilityrate and acrosome intact rate after the dog seminal fluid freezes can be significantly improved.

The invention discloses a semen protection solution for improving the quality of frozen semen as well as a preparation method and application of the semen protection solution. The semen protection liquid contains active ingredients, and the active ingredients are prepared from rhodiola rosea polysaccharide and astragalus polysaccharide. The semen protection liquid has the effects of resisting oxidation, improving the survival rate of frozen sperms and the like, can be effectively used for semen freezing protection, can effectively improve the plasma membrane integrity rate, the sperm motility, the DNA integrity rate, the acrosome integrity rate and the like of unfrozen sperms, and improves the quality of the frozen sperms. The method has the advantages of wide raw material source, safety, environmental protection, simple preparation process, low production cost and the like.

The invention discloses application of taxifolin and kaempferol in preparation of a livestock cryopreservation agent. Meanwhile, the invention discloses a livestock cryopreservation agent containing taxifolin, kaempferol and a freezing base solution. According to the invention, taxifolin and kaempferol are combined for cryopreservation of semen of pigs, sheep and cattle for the first time; the application is reliable in effect, easy to popularize and suitable for artificial insemination of pig, sheep and cattle tubule frozen semen, long-term storage of the semen after ultralow-temperature freezing and semen deposition after long-distance transportation, the sperm motility rate, acrosome integrity rate and plasma membrane integrity rate of the frozen-thawed pig, sheep and cattle can be remarkably increased, and the fertilization capacity of sperms is kept.

The invention relates to dairy goat frozen semen diluent, its preparation method and dilution method, and a preparation method of dairy goat frozen semen thin tube frozen semen. In the existing frozen semen preservation method, the plasma membrane structure of the sperm is changed due to the impact of low temperature, which seriously affects the integrity of the plasma membrane and its viability and fertilization ability after recovery. The present invention performs two-step dilution, balance and two-step cooling thin tube freezing treatment on fresh semen, wherein, the basic diluent is composed of Tris, citric acid, glucose, trehalose, Vc, GSH, BSA, ATP, compound antibiotics, egg yolk and water The frozen diluent is made up of basic diluent plus glycerin; after the diluent is used to dilute and balance the fresh semen, it is divided into thin tubes and stored in liquid nitrogen. The invention has good effect on cryopreservation and cryopreservation of dairy goat semen, effectively improves the survival rate after thawing, the integrity rate of acrosome and the integrity of plasma membrane, reduces the deformity rate, and improves the production efficiency and quality of frozen semen of dairy goats.

The invention relates to an animal semen diluting powder and application thereof, and belongs to the technical field of animal reproduction. The animal semen diluting powder is dissolved in a PBS buffer solution to prepare a pig semen diluting solution; the semen diluting solution comprises 0.1-0.3 g / L of liquiritin, 1-3 g of sodium glutamate, 0.5-1.0 g of L-arginine, 0.1-0.3 g of N-acetylcysteine, 1-3 g of fructose, 10-15 g of glucose, 2-4 g of potassium citrate, 0.5-1 g of potassium chloride, 1-2 g of EDTA, 1-2 g of sodium carbonate and 0.1-0.3 g of origanum oil. The liquiritin can also be replaced with glycyrrhetinic acid. The animal semen diluting powder does not adopt any antibiotics, and various components have a synergistic effect, so that the time and the motility rate of preservedsperms can be obviously improved, the integrity rate of acrosomes can be improved, and the pregnancy rate of artificial insemination after dilution can be increased.

The invention relates to a yak frozen semen diluent and a preparation method thereof, and belongs to the technical field of bioengineering. The frozen semen diluent is prepared from a herba cistancheextracting solution, a rhizoma curculiginis extracting solution, a semen euryales solution, sodium citrate, glucose, raffinose, resveratrol, tetrasodium glutamate diacetate, corn oligopeptide and thelike. The herba cistanche extracting solution, the rhizoma curculiginis extracting solution and the semen euryales liquid are introduced to maintain sperm activity, resveratrol is used as an antioxidant, tetrasodium glutamate diacetate is used as a chelating agent, raffinose is used for guaranteeing the cell membrane protection function, and the prepared yak frozen semen diluent can obviously improve the post-freezing motility rate and the acrosome integrity rate of sperms and improve the post-freezing quality; moreover, due to the fact that no animal-derived component exists, the risk of infection or diseases is avoided, and application and popularization are easy.

The invention belongs to the technical field of sheep breeding, and relates to a sheep semen diluent which comprises the following components in parts by mass: 1-2 parts of citric acid, 1.5-2.5 parts of fructose, 2-4 parts of Tris, 5-18 parts of yolk, 0.1-0.4 part of penicillin sodium, 0.1-0.4 part of streptomycin sulfate and 100 parts of ultrapure water. According to the invention, the preservation time of the sheep semen is effectively prolonged, slow and stable reduction of sperm movement performance is maintained, and damage to sperm structures such as plasma membranes and acrosomes is slowed down. The invention provides the diluent suitable for sheep semen preservation, and the semen preservation quality is remarkably improved.

The invention discloses a composition for promoting fertilization of frozen semen of a mouse. 0.01 mg / ml PVA and 0.75 mM methyl-beta-cyclic dextrin (MBCD) are added to an m HTF solution to serve as asperm capacitation solution; and 1.0 mM reduced glutathione is added to an m HTF solution to serve as a sperm in-vitro fertilization solution. The sperm capacitation solution used in the invention canremarkably protect the integrity of a plasma membrane and an acrosome, reduce the influence of ROS, and prominently improve survival rate and mobility of a sperm; and in combination with the in-vitrofertilization solution containing the reduced glutathione in the invention, an in-vitro fertilization success rate of a frozen-thawed sperm of a genetically modified mouse with an inbred strain C57BL / 6J as a background can be improved remarkably, and a two-cell developmental rate after fertilization is 60-65%.