61 results about "Sperm freezing" patented technology

A practical method for freeze storage of fish sperm features that the diluent, antifreezing agent and container for the freeze storage of fish sperm chosen from many schemes, and a three-step freezing mode and a quick thawing method are designed.

Described herein is a composition that comprises a cryoprotectant; a membrane protectant that stabilizes or assists in stabilization of membranes of sperm; and a free radical scavenger (e.g., a reducing agent, an antioxidant).

The invention relates to an extremely-low sperm freezing protective agent and application thereof. The protective agent is prepared from the following ingredients: sodium chloride, potassium chloride, magnesium sulfate, monopotassium phosphate, calcium chloride, fructose, trehalose, sodium citrate, glycin, glutamine, pentoxifylline, vitamin E, HAS (human serum albumin), glycerin and gentamicin sulphate. The extremely-low sperm freezing protective agent can protect less sperms with low activity and without forward activity ability in seminal fluid, can remarkably improve the activity ability of the sperms which swing weakly and cannot move forwards in seminal fluid after freezing and thawing, has no obvious influence on the physiological function of the sperms, has high freezing anabiosis rate, can be used for cryopreservation of less and weak sperms, testicle sperms and epididymis sperms, and contributes to the successful development of assisted reproductive technology.

The invention relates to a method for freezing and storing diluted semen of cow. Wherein, the invention is characterized in that: preparing said diluted semen into A, B, C, D deals; mixing them with right ratio; adding 5-20% pig semen into C and D deals, to improve the semen acrosome integral rate of cow semen control liquid, and reduce the density of semen, to save cost. The experiment has proved that the semen survival rate of stored diluted semen can reach 50.88-62.88%, while the average number is 57.41%, which is 15.13% higher than traditional method, and the semen acrosome integral rate can reach 50.88-53.20%, while the average number is 51.90%, which is 7.0% higher than traditional method.

The invention discloses a method for inducing cleavage gynogenesis of fish fry of cynoglossus semilaevis, which comprises the steps of cynoglossus semilaevis sperm freezing storage, ultraviolet radiation of sperms, and induction and identification of the cleavage gynogenesis of the fish fry. The cynoglossus semilaevis sperm freezing storage method is established, and a method for inhibiting the first cleavage to induce the multiplication of embryo chromosome of the gynogenesis of the cynoglossus semilaevis through hydrostatic pressure is adopted so as to screen effective dilution and freezing method for the cynoglossus semilaevis sperm freezing storage and screen the hydrostatic pressure shock starting time, hydrostatic pressure and processing time suitable for the cleavage gynogenesis of cynoglossus semilaevis; and an identification method for the gynogenesis of the fish fry of the cynoglossus semilaevis is established. In the method, the fish fry of the cynoglossus semilaevis through cleavage gynogenesis is obtained for the first time, and the inductivity of the gynogenesis of the fish fry reaches 0.25 percent; and the method has the characteristics of advancement, high efficiency, reliability and reliability, can be used for solving the problems that male cynoglossus semilaevis grows slowly, the ratio of female cynoglossus semilaevis is low and the culture cost is high, and has significant application value and wide promotion and application prospect in the aspects of breeding of cynoglossus semilaevis fry, sex control, culture of all female fry, pure line breeding and the like.

The invention discloses a trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof. The trace sperm cryopreservation carrier is in a rectangular, square or circular horizontal or groove shape and is made of a transparent plastic or transparent glass, and the size of the trace sperm cryopreservation carrier can bear 1-3 0.1-10.0 mu L freezing liquid micro drops. The freezing and thawing method comprises the following steps: after preparing the freezing liquid micro drops on the trace sperm cryopreservation carrier, inserting the trace sperm cryopreservation carrier into a sperm culture dish covered with oil, grabbing sperms by a microinjection needle, putting into the freezing liquid micro drops, taking out the cryopreservation carrier, soaking up mineral oil by virtue of an absorbent paper, rapidly freezing, directly inserting the cryopreservation carrier into an ICSI vessel when thawing, and thawing the sperms by utilizing the mineral oil with the temperature of 37 DEG C. According to the trace sperm cryopreservation carrier and the corresponding freezing and thawing method thereof disclosed by the invention, as the cryopreservation carrier is inserted into the sperm culture dish covered with oil, and the sperms are grabbed and frozen, a special moisture retention device is not needed, and evaporation of the freezing liquid micro drops is not needed to be worried about; the operation is very simple, the flexibility and reliability of assisted reproductive treatment for a patient with azoospermia are greatly improved, the sperm recovery rate can reach 100 percent, and the sperm viability is about 60 percent.

The invention relates to a selection and culture method of excellent breed large-tooth flounder featuring fast growth and high survival rate. The technical method comprises: culture and reproduction regulation and control of large-tooth flounder breeding group, establishment of family by sperm freezing technology, culture and standardization of the family, determination and analysis of number and properties, estimation of genetic parameters, screening of excellent family, and establishment of excellent breed by inducing thelykaryon of large-tooth flounder to develop by using heterologous frozen sperms. The invention can select and culture new breed of cultured large-tooth flounder featuring fast growth and high survival rate, screen out four excellent large-tooth flounder families featuring fast growth and high survival rate (0703, 0719, 0750 and 0751), and establish seven excellent breeds of large-tooth flounder developed from thelykaryon (0920, 0921, 0927, 0938, 0939, 0946 and 0947). The technical method can be applied to new variety culture of large-tooth flounder cultivation, solve the problems of culturing large-tooth flounder fish in sea water such as slow growth and low survival rate, and enhances economic benefit of large-tooth flounder cultivation.

The invention relates to a sperm cryoprotectant and a preparation method thereof. Every 125 ml of the sperm cryoprotectant is prepared from saccharose, sodium bicarbonate, glycerol, human serum albumin and water obtained by distilling for five times. The novel sperm cryoprotectant does not contain yolk and only contains saccharose, sodium bicarbonate, glycerol, 20% (g / mL) human serum albumin and other commonly-used reagents with definite chemical components, the risk of animal source pathogenic bacterium infection does not exist, the acquired freezing effect is similar to that of an existing formulation, and the average value of the sperm freezing-thawing rate is about 75% to 80%.

The invention provides cryopreservation solution of horse sperms and a freezing method thereof. The freezing method comprises the steps of firstly, applying mare milk and yolk low-density lipoprotein after special treatment to the cryopreservation solution of horse sperms at the same time; adding the cryopreservation solution by sub-steps in the freezing process; and controlling cooling, lower liquid nitrogen fumigation height and shorter fumigation time. The quality of freezing the horse sperms is obviously improved by the technology; the artificial insemination efficiency of frozen semen of the horse is greatly improved.

The invention discloses an inbreeding coefficient calculation model-based dairy cow hybridization method. The method comprises the following steps of: recording information of each dairy cow through abreeding bull electronic document database and a breeding cow electronic document database, wherein a database table structure comprises detailed information such as breeding bull numbers, breeding bull names, fathers, mothers, families, strains, dates of birth, places of birth, bodily forms, TPI and performance indexes; and when pairing is started, implanting the numbers of to-be-paired dairy cows and the numbers of sperm freezing bulls by an information system, automatically pairing and combining the sperm freezing parameters of each cow and each bull one by one by a computer system, reading a family tree relationship of the paired dairy cows in each group of pairing, constructing an electronic family tree graph, and processing data by utilizing an inbreeding coefficient calculation model so as to obtain an inbreeding coefficient of the combination. According to the method, systematic processing of large data volume can be rapidly and correctly carried out, so that unnecessary humanlabor can be greatly decreased, the correctness of calculated values can be ensured and the processing speed can be improved.

The invention relates to the technical field of assisted reproduction, and discloses a cryoprotectant and an application thereof, a sperm refrigerating fluid and a preparation method of the sperm refrigerating fluid. The sperm refrigerating fluid comprises a cryoprotectant, a composition and a basic culture solution, wherein the cryoprotectant comprises 0.3 to 1.1 mg / L of paclitaxel, 16 to 22 g / Lof trehalose, 7 to 13 g / L of cane sugar and 80 to 250 mL / L of glycerinum; the composition comprises 12 to 26 mg / L of resveratrol, 0.2 to 1.9 mg / L of reduced glutathione, 436 to 775 mg / L of lecithin, 0.5 to 1.6 mg / L of cholesterol, 9 to 17 mg / L of ATP sodium salt, 11 to 28 mg / L of heparin, 93 to 299 mg / L of phosphatidylserine and 5 to 25 mg / L of coenzyme Q10. The sperm refrigerating fluid can reduce damage of sperms in low-temperature freezing and freezing resuscitation processes, improve vitality of the sperms after freezing resuscitation, prolong survival time of the sperms after freezing resuscitation, promote sperm capacitation and enhance penetrating fertilization capacity of the sperms to egg cells. Thus, the fertilization success rate and blastocyst formation rate of the ova are improved, and finally the effect of improving the pregnancy success rate of the in-vitro fertilization assisted reproduction technology is achieved.

The invention provides a human sperm cryopreservation liquid, a human sperm cryopreservation method, and a human sperm cryopreservation and recovery method. The cryopreservation liquid comprises a yolk liquid, glycerol, antibiotics, a refrigeration buffer solution, and quercetin or derivative thereof. The human sperm cryopreservation and recovery method comprises the steps of using the human sperm cryopreservation liquid, performing cold balance at a temperature of 4 DEG C, and then performing refrigeration at a temperature of minus 80 DEG C, and next, transferring to a liquid nitrogen tank for cryopreservation; and performing unfreezing and recovering in a water bath box at a temperature of 37 DEG C before use. The sperm cryopreservation liquid, added with the quercetin at a proper concentration, can effectively protect sperms and can remarkably improve survival rate and motility of revived sperms in the subsequent sperm recovery process, so that occurrence of sperm ROS and apoptosis can be reduced; and therefore, the human sperm cryopreservation liquid has application potential in the assisted reproductive technology.

The invention relates to a sperm freezing fluid and a preparation method thereof and belongs to the technical field of human assisted reproduction. The sperm freezing fluid is prepared by dissolving glycerinum into a salt solution with cane sugar, antibiotics and an indication agent in a concentration of 20-30%. Due to addition of the glycerinum and the cane sugar in the sperm freezing fluid provided by the invention, the glycerinum and the cane sugar are adopted as cell protection agents which can not only gently and effectively protect activity of sperms and completeness of DNA (Deoxyribonucleic Acid), but also protect the activity of testicular tissue cells, furthermore the activity of sperms in testicular testes is protected, the fertility rate of the sperms is increased, and the security of assisted reproductive technologies is powerfully ensured.

The invention discloses a sperm cryoprotectant for a C57BL / 6J mouse and a genetically engineered mouse taking the C57BL / 6J mouse as genetic background. The components of the cryoprotectant comprise raffinose, skim milk, and yolk, wherein the volume ratio of the yolk is between 10 and 30 percent. The mouse sperm cryoprotectants is added with egg yolk liquid so that sperms of the C57BL / 6J mouse and the in vitro fertilization rate of the genetically engineered mouse taking the C57BL / 6J mouse as the genetic background are increased by 4.5 times maximally compared with that of the cryoprotectant purely using the raffinose and the skim milk, thus the fertilization rate is effectively improved.

The invention relates to a preferential sperm freezing culture device which comprises freezing slide glass, a sperm tank, a sperm swimming channel, a collecting tank and a blocking piece. The volume of the sperm tank ranges from 50 microliters to 500 microliters, the volume of the collecting tank ranges from 5 microliters to 20 microliters, the width of the sperm swimming channel ranges from 10 micrometers to 30 micrometers, and the sperm swimming channel is connected with the sperm tank and the collecting tank. The preferential sperm freezing culture device has the advantages that the preferential sperm freezing culture device is applicable to screening preservation of frozen sperm, motile sperm can be automatically separated by the design of the sperm tank, the sperm swimming channel andthe collecting tank, operation amount is decreased, mechanical damage to the sperm caused by manual picking is avoided, the proportion of usable motile sperm is increased, the freezing slide glass ofall existing sperm can be completely replaced, the preferential sperm freezing culture device can be applied to the fields of normal sperm optimization sperm periodic IVF (in-vitro fertilization), ICSI (intracytoplasmic sperm injection) or artificial insemination, sperm collection of oligospermia and asthenospermia patients, surgical sperm extraction of azoospermatism patients, sperm collection and the like, and has a wide popularization and application prospect.

The invention discloses a method for freezing the semen of a Macaca fascicularis. In the method, fresh semen activity of a semen sample is required to be more than or equal to 0.7. The freezing method comprises the following operating steps of: 1) freezing diluent prescription and diluent preparation for the semen of the Macaca fascicularis; 2) semen dilution; 3) semen package; 4) semen balance; 5) semen freezing; and 6) frozen semen unfreezing. The method has the following advantages that: by using the technology of the invention, in the processes of freezing and unfreezing the semen of the Macaca fascicularis, the motility rate and the recovery rate of the semen after freezing and unfreezing respectively reach 45 percent and 62 percent, so that the reproductive efficiency of good male Macaca fascicularis can be brought into full play, genetic gene of good Macaca fascicularis can be conserved, and referential experience is provided for the research on semen cryopreservation of other rare or endangered primates at the same time.

The invention relates to the field of sperm freezing, and particularly relates to a sperm freezing medium as well as a preparation method and application thereof. The sperm freezing medium comprises the following components: potassium chloride, magnesium sulfate, calcium chloride, sodium bicarbonate, dipotassium phosphate, sodium pyruvate, calcium lactate, glucose, saccharose, glutamine, glycine,HEPES, gentamicin, glycerinum, human serum albumin, vitamin E, vitamin C and vitamin B12. According to the sperm freezing medium, all components are matched with each other to comprehensively maintainthe osmotic pressure, the straining capacity of sperm to resist the outside environment change; the freezing medium is used for storing sperm in low temperature; the sperm survive rate and the activity can be maintained for a long term.

A case of goose seminal fluid chill diluent and application method thereof belongs to animal breed technique field, the reagent case including: component I: sodium glutamate 0.4-0.6, potassium dihydrogen phosphate 1.4-2.2, dipotassium hydrogen phosphate 0.2-0.5, homocycteine 0.0006-0.01, fructose 0.9-1.2, inositol 0.04-0.08, raffinose 0.5-0.8, sodium oxacillin 8-12, streptomycin sulphate 0.07-0.15, calcium chloride 0.01-0.02, 3-carboxymethylaminomethane 0.1-0.3; component II: double distilled water 100; component III: protamini sulfas 0.04-0.06 g; component IV: DMA 4.5-5.5, glycol: 4.5-5.5 ml. Useful effects: using components of 3-carboxymethylaminomethane and protamini sulfas and quantifying accurately. Compounding each component of goose seminal fluid chill diluent as packing type of case firstly. The method is simple and easy to operate, and improve applying effects of the case.

The invention relates to a sperm freezing method which mainly comprises the following steps: 1) diluting and well mixing sperm and freezing liquid according to a ratio of 1:1; 2) slowly cooling at 4 DEG C, after 3 hours, individually adding the mixture in plastic tubules of 0.25-0.35 ml and sealing, putting on a suspended object with a distance from a liquid nitrogen surface of 4-6 cm at a temperature of from -90 to -95 DEG C; 3) putting the tubules into the liquid nitrogen after 10-15 minutes. The sperm freezing method provided by the invention has simple operations, and the frozen-thawed sperm still has high fertilization ability and an high embryonic development rate.

The invention belongs to the technical field of sperm freezing, and particularly relates to a Hexagrammos otakii sperm productive cryopreservation method. The method comprises the following steps: after collecting sperms, uniformly mixing seminal fluid with a cryoprotectant, and pre-cooling in a refrigerator at 4 DEG C for 5 minutes; freezing a straw at -20 DEG C for 10 minutes, putting the straw into a straw sleeve, and putting the straw sleeve into an aluminum cryopreservation strip; putting the cryopreservation strip into a pail of a liquid nitrogen tank, putting the pail on the liquid nitrogen surface of the liquid nitrogen tank, cooling for 10 minutes under the condition of -160 DEG C to -180 DEG C, and then putting the cryopreservation strip linto liquid nitrogen for preservation; wherein the cryopreservation liquid is prepared by taking boiled and cooled sterilized natural seawater as a diluent and ethylene glycol with the final concentration of 15% (v / v) as a protective agent. Based on production requirements, the invention provides a simple and batch productive sperm cryopreservation method, and provides technical support for selective breeding of improved varieties of hexagrammos otakii and fishery production; according to the method, the sperm anti-freezing liquid is simple in proportioning, the freezing procedure is relatively convenient and fast, and the requirement for cryopreservation of productive sperms can be met.

The invention discloses an automatic sperm cold storage table and controller, which comprises a bottom plate, wherein a guide rail is arranged on the bottom plate; an object carrying table is arrangedon the guide rail and is connected with a power mechanism driving the object carrying table to move along the guide rail; the power mechanism is connected with a controller, and comprises a motor anda screw; the object carrying table is provided with a threaded hole; the lower end of the threaded hole is movably connected with the bottom plate; the motor is arranged at the lower side of the bottom plate; the screw rod is connected with the threaded hole through screw threads; a control base is arranged at the lower side of the bottom plate; the controller and the motor are arranged in the control base; after the carrying of a sperm freezing sheet, the object carrying table can automatically and slowly descend into liquid nitrogen; the operation is convenient; the liquid nitrogen suffocation on the sperm freezing sheet clamped by professional medical personnel is not needed; the energy consumption of medical personnel is greatly reduced. In addition, a plurality of sperm freezing sheets can be carried and placed onto the object carrying table; the batched sperm liquid nitrogen suffocation freezing can be performed at the same time; very high cold storage efficiency is realized.

The invention relates to a sperm freezing device which comprises a tank body, a liquid level adjusting mechanism, a bracket, a timer and a controller, wherein an accommodation cavity is formed in thetank body; the accommodation cavity is internally provided with a partition plate for partitioning the accommodation cavity into a standby storage cavity and a freezing cavity; the inner wall of the freezing cavity is provided with a first position and a second position at an interval; the liquid level adjusting mechanism is mounted on the partition plate and is used for communicating the standbystorage cavity with the freezing cavity when the liquid level in the freezing cavity is lower than a preset value, and thus the liquid level in the freezing cavity is kept constant; the bracket is mounted on the inner wall of the freezing cavity in a sliding manner along a connection line direction of the first position and the second position; the bracket is used for holding sample cryopreservation tubes with sperm samples; the timer is used for setting fumigation time according to preset instructions; and the controller is electrically connected with the timer, and is used for controlling the bracket to move to the second position from the first position when the timer meets the fumigation time. Therefore, due to adoption of the sperm freezing device, the quality of frozen sperms is high.

The invention discloses a rat sperm cryopreservation agent (RS-CPA) for a SD rat and a gene engineering rat under a SD rat genetic background. The RS-CPA comprises 5% of beam milk extract as a RS-CPA main component and also comprises raffinose. Through use of the beam milk extract, the RS-CPA greatly improves resurgent sperm vitality and a fertilization rate and realizes a refrigeration recovery rate of 10% or more.

The invention provides cryopreservation solution of horse sperms and a freezing method thereof. The freezing method comprises the steps of firstly, applying mare milk and yolk low-density lipoprotein after special treatment to the cryopreservation solution of horse sperms at the same time; adding the cryopreservation solution by sub-steps in the freezing process; and controlling cooling, lower liquid nitrogen fumigation height and shorter fumigation time. The quality of freezing the horse sperms is obviously improved by the technology; the artificial insemination efficiency of frozen semen of the horse is greatly improved.

The invention discloses an ultralow-temperature cryopreservation method for Yellow River carp sperms. The method specifically comprises the following steps: preparing a sperm preserving fluid from anantifreeze agent methanol and a diluent; mixing the Yellow River carp seminal fluid and the sperm preserving fluid according to a volume ratio of 1: 5 in a cryopreservation tube, wherein the volume concentration of the antifreeze agent methanol in the mixed solution of the Yellow River carp semen and the sperm preservation solution is 17%; and then pre-cooling for 5 minutes at the temperature of 4DEG C, balancing for 12 minutes at the position 5 cm above the liquid nitrogen, putting into the liquid nitrogen and carrying out ultralow-temperature treatment for 1 day, whrein the diluent is prepared from the following raw materials in percentage by weight: 0.95 of NaCl, 0.05% of KCl, 1.5% of glucose and the balance of water. The ultralow-temperature cryopreservation technology for the YellowRiver carps is established, a technical guarantee is provided for establishment of a Yellow River carp sperm freezer and the technical support is provided for germplasm preservation and genetic breeding work of the Yellow River carps.

The invention discloses a trace sperm freezing and thawing method and an automatic clamping apparatus thereof. The method comprises: S1, freezing sperms; and S2, thawing the sperms, wherein a fixing frame is placed on the bottom portion of the inner side of a liquid nitrogen box, a lifting motor is fixed on the top portion of the fixing frame, and the output shaft of the lifting motor is fixedly connected to a lifting screw rod penetrating through the fixing frame. According to the present invention, the apparatus can simultaneously freeze a plurality of freezing carrying rods, such that the operation is convenient, efficient and fast, and the sperm micro-droplets on the freezing carrying rods are subjected to uniform and accurate fumigation by the condenser so as o substantially improve the freezing resuscitation rate of the trace sperms and reduce the mistakes and the frostbite of the operator; and with the method and the apparatus, the problems that the sperm recovery is easy to fail after the sperms of paints with oligospermatism and asthenospermia, the testis sperms and the epididymal sperms are frozen in the current sperm freezing technology is well solved so as to substantially improve the assisted reproductive therapy flexibility on patients with azoospermia.

The invention relates to a cryogenic diluent of the semen of Tibetan mastiffs, which belongs to the technical field of animal reproduction and comprises 1 fluid and 2 fluid; wherein, the 1 fluid is prepared by the following steps: the raw materials comprise 0.20 to 0.50g of calcium chloride, 0.18 to 0.36g of potassium chloride, 0.22 to 0.45g of magnesium chloride, 3.5 to 5.85g of sodium chloride, 0.30 to 0.55g of monosodium phosphate, 1.38 to 3.20g of sodium bicarbonate and 0.025 to 0.036g of sodium pyruvate, and double distilled water is added to obtain constant volume of 1L; the 2 fluid is prepared by the following steps: 35 to 55ml of the 1 fluid, 20 to 30ml of calf serum, 20 to 25ml of yolk, 5 to 10ml of glycerin and 1000IU / ml of penicillin are mixed to prepare to the diluent with the penetrating pressure of 300 to 330mosmo / l and the pH value of 6.5 to 7.0; the diluent leads the activity of the cryogenic diluent of the semen of Tibetan mastiffs to reach 0.6 to 0.7, and the conception rate and the average number of litter size to respectively reach 77.3 percent to 84 percent and four to five so as to reach or near the selfing level, thus being a cryogenic diluent of the semen of Tibetan mastiff with practical application and promotion value.

The invention relates to the technical field of sperm freezing, in particular to a single sperm freezing carrier in combination with monoclonal antibodies and a freezing and unfreezing method. Firstly, an APTES is utilized to make the outer surface of a slide become a carbanyl group, the carbanyl group very easily reacts with an amino group on the surface of immunoglobulin, then some monoclonal antibodies with a certain characteristic combine with the surface specificity of the sperm, and the adhesive force of the antibodies fixes the sperm to the surface of the slide. The invention provides amethod capable of fixing one or more sperms to the surface of the carrier, and the sperms can be prevented from being lost in the storage, transportation and operation process.

The invention discloses sperm refrigerating fluid for human assisted reproduction and a preparation method thereof. The cane sugar in the sperm refrigerating fluid is replaced trehalose. The sperm refrigerating fluid comprises 95.00-105.00 mmol / L of sodium chloride, 3.45-6.55 mmol / L of potassium chloride, 0.55-0.65 mmol / L of magnesium sulfate, 1.75-2.22 mmol / L of calcium chloride, 0.18-0.39 mmol / Lof sodium dihydrogen phosphate, 0.58-1.78 mmol / L of sodium citrate, 3.85-5.45mmol / L of sodium bicarbonate, 15.00-25.00mmol / L of HEPES and MOPS, 0.38-0.68 mmol / L of glucose, 0.28-0.42 mmol / L of sodium pyruvate, 12.48-23.68 mmol / L of sodium lactate, 5-15 mg / mL of human serum albumin, 7-14 [mu]g / mL of gentamicin sulfate, 25-35% (v / v) of glycerin, and 0.15-0.65 mol / L of trehalose. The preparation method comprises the following steps: weighing various components, dissolving the components in injection-grade water, performing filtering and sterilizing by using a 0.2 [mu]m filter membrane, performing sampling and testing, wherein the pH value is controlled to be 7.00-7.40, the osmotic pressure ranges from 1720 mOsm / Kg to 2020 mOsm / Kg, the bacterial endotoxin is less than 0.25 EU / ml, and the blastocyst formation rate is greater than or equal to 80% when 1 cell mouse embryo is cultured for 96h.