59 results about "Sperm freezing" patented technology

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 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 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 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 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 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.

The invention discloses a sperm cryopreservation method based on a protective agent, which comprises the following steps: (a) diluting a sperm sample by using a semen-rare sperm cryoprotectant, the volume ratio of the semen-rare sperm cryoprotectant to the sperm being 1: (1-2); (b) placing the semen-rare sperm cryoprotectant mixture at room temperature to act for at least 5 minutes; (c) putting the semen-rare sperm cryoprotectant mixture into a preservation container, buckling the container on a bracket, and then carrying out fumigating on a liquid nitrogen surface for 10 minutes; and (d) transferring the bracket loaded with the preservation container into a liquid nitrogen storage tank at -196 DEG C for storage. The liquid nitrogen storage tank in the step (d) comprises a tank body, a first vacuum layer, supporting legs, a freezing bin, a mounting opening, a freezing frame, a safety fastener, a lifting device, a clamping device and a two-axis coordinate positioning device. The freezing frame is arranged, so that more frozen test tubes can be stored in order. The two-axis coordinate positioning device is arranged, so that code storage and positioning rapid extraction can be realized, and the efficiency is improved.