32 results about "External fertilization" patented technology

The invention relates to the technical field of livestock and poultry breeding, in particular to a goat hybridization breeding method capable of achieving complete-cycle nutrition enhancement. The core of the method is that systematic nutrition intervention runs through the whole breeding process. Before hybridization, performing differential nutrition pretreatment on the parents for 60 days: feeding the female parents with a breeding nutrition enhancer added with components such as D-chiro-inositol and N-acetylcysteine, and supplementing sperm motility enhancer containing acetylated L-carnitine and ergothioneine to the male parents. In the gamete treatment stage, the oocytes are subjected to in-vitro maturation culture in a special culture solution containing components such as D-chiro-inositol and a growth differentiation factor 9; after the sperms are subjected to gradient centrifugal screening, completing in-vitro fertilization in an optimized fertilization culture solution; after the embryos develop into blastocysts in the sequential culture system, carrying out laparoscopic transplantation, and carrying out staged precise nutrition culture on the pregnant ewes. According to the method, through multi-level nutrition enhancement, the reproductive capacity of the goats and the growth performance of offspring are effectively improved.

A method of artificial-intelligence-based robotic pipetting for spermatozoa preparation includes positioning a vessel containing a semen sample within a staging mechanism. The method includes using a robotic pipettor to make at least two droplets within a dish on the staging mechanism. The method includes using the robotic pipettor to connect the at least two droplets with a medium channel according to a programmable design commanded by an artificial intelligence / machine learning system (AI / ML system). The method includes using the robotic pipettor to deposit a quantity of sperm from the semen sample into one of the droplets. The method includes using the AI / ML system to optically scan the quantity of sperm to produce first and second image objects using an imaging system that includes a microscopy system, a camera system, and a lighting system. Creation of the image objects is separated by a specified time duration.

The invention discloses an artificial breeding method of lota lota, which comprises the following steps: S1, selecting robust lota lota as parent fish, placing the parent fish in an indoor culture tank for temporary culture, and inducing gonad to develop mature by regulating and controlling water temperature; s2, performing artificial spawning induction on the parent fish with mature gonad to obtain ova and semen, and performing in-vitro fertilization on the ova and the semen to obtain fertilized ova; s3, performing two-step soaking treatment on the fertilized eggs by using a composite biological functional liquid containing a liquid A and a liquid B, and then transferring the fertilized eggs into a hatching facility for hatching to obtain fish fries; and S4, breeding the hatched fish fries. According to the artificial breeding method of the lota lota, the survival rate of fertilized eggs and the survival rate in the fry breeding stage are increased to a high level, large-scale and controllable artificial breeding of the lota lota becomes possible, and the overall technical scheme has high stability and industrial application value.

The invention belongs to the technical field of biology, and particularly relates to an acrossocheilus fasciatus sperm cryopreservation method which comprises the following steps: constructing a simulated water solution for acrossocheilus fasciatus in-vitro fertilization, collecting fresh sperms, detecting the survival rate, preparing a mixed solution of the sperms and anti-freezing components, sub-packaging, cooling and freezing and the like. According to the method, simulated water quality for in-vitro fertilization of acrossocheilus longifin is constructed, the survival rate of sperms observed and detected in the environment is closer to the fertilization condition during thawing activation in the future, and then the sperms with the higher fertilization success rate after thawing are screened out; meanwhile, the constructed simulated water solution can be used as a part of a cryopreservation solution, so that the remaining anti-freezing components can be directly added into the screened sperms, a mixed solution of the sperms and the anti-freezing components can be rapidly prepared under the condition that the service life of the sperms is not remarkably shortened, and the mixed solution is subpackaged, cooled and frozen; the improved anti-freezing component contains trehalose, soybean phospholipid and tea polyphenol, so that the survival rate and activity of the acrossocheilus fasciatus sperms after cryopreservation can be effectively improved.

A method for automated ICSI includes providing an egg, using AI to position a robotic ICSI needle, moving the robotic ICSI needle forward into the egg, stopping the robotic ICSI needle at the end point of an injection path, breaking the egg membrane (oolemma) using a piezoelectric pulse, and depositing the sperm in the egg. The methods can also include confirming that the sperm is out of the needle, moving the needle out of the egg, and releasing the egg from a holding pipette.

The utility model discloses a movable embryo laboratory which comprises a movable chassis and a laboratory box body, the laboratory box body is arranged above the movable chassis, and a traction device is fixedly arranged at the front end of the walking direction of the movable chassis. A unit operation and maintenance room, a disinfection room, an ovum collection room, an in-vitro fertilization culture room and a cryopreservation room are arranged in the laboratory box body from front to back in the walking direction of the movable chassis, and every two adjacent areas of the ovum collection room, the in-vitro fertilization culture room and the cryopreservation room are communicated through doors. Wall bodies of the disinfection room, the ovum collection room, the in-vitro fertilization culture room and the cryopreservation room are provided with delivery windows for communicating two adjacent areas, and the disinfection room is partitioned in the front-back direction to form a gas cylinder room and a decontamination room. According to the utility model, sperm and egg sample treatment time can be shortened, and embryo culture success rate is improved; sample loss caused by long-distance transportation is avoided; the operation flexibility is improved, and the embryo treatment cost is reduced; the method is convenient to deploy quickly and is suitable for pastures under different terrains and climate conditions.

The present disclosure relates to a method for constructing a mouse model with short telomere. Specifically, the present disclosure obtains fertilized eggs by fertilizing the sperms and eggs of mice in vitro, and the fertilized eggs are cultured to the blastocyst stage in vitro and then transferred into the surrogate female mouse for development, thereby producing mice with short telomere. The method of the present disclosure does not require gene editing, has a short modeling cycle, and the effect is reliable and stable. By changing only the environment during embryo transfer, the telomere elongation process in the embryo is interfered, so as to successfully construct a progeny model with short telomere. There is no significant effect on the reproductive rate of female mice. Therefore, the present disclosure can provide a method for constructing a mouse model with short telomere for exploring the mechanism of telomere shortening and studying telomere-related phenotypes such as aging.

The application discloses a kind of water and fertilizer integrated machine's fertilization diagnostic system and method, system includes: fertilizer storage barrel, fertilizer storage barrel is used to store fertilizer solution in;Fertilizer barrel, fertilizer barrel is communicated with fertilizer storage barrel by first connecting pipeline, to fertilizer storage barrel and fertilizer barrel are configured to be connected device structure, the radius of fertilizer barrel is less than the radius of fertilizer storage barrel, fertilizer barrel is connected with second connecting pipeline in the position close to bottom, to be used for external fertilization;Liquid level sensing switch sensor, liquid level sensing switch sensor is arranged on the outer wall of fertilizer barrel, and liquid level sensing switch sensor is movable along the outer wall of fertilizer barrel, to detect the fertilizer solution in fertilizer barrel;Movable assembly, movable assembly is connected with liquid level sensing switch sensor, to drive liquid level sensing switch sensor movable in vertical direction along fertilizer barrel;Control box, control unit is arranged in control box, liquid level sensing switch sensor is connected with control unit.

The invention discloses a high-efficiency C57BL / 6 mouse in-vitro fertilization method, relates to use of an in-vitro fertilization method in the technical field of biology, and particularly relates to a method for obtaining more high-quality fertilized eggs for gene editing in an in-vitro fertilization process of small animal gene editing model construction. Therefore, the animal cost of gene editing is greatly reduced, and wide application is promoted.

The present invention relates to transgenic ornamental barbs, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic barbs and methods of providing them to the ornamental fish industry for the purpose of marketing.

A system and method for improving sperm selection during in vitro fertilization (IVF) procedures includes a sperm containment apparatus equipped with a capture system designed to isolate motile sperm as they respond to the presence of a retrieval instrument within the sperm solution. As more motile and potentially healthier sperm move away from the retrieval instrument, they are selectively corralled within certain regions of the apparatus. The capture mechanism may include various structures, such as a barrier member including one or more passageways which direct sperm movement. By isolating sperm based on their motility, this invention enhances the selection process, improving the chances of successful fertilization with higher-quality sperm. This system provides a more efficient and targeted approach to sperm retrieval compared to conventional methods.

To provide a method for producing a selected sperm, capable of improving fertilization rate in in-vitro fertilization or artificial fertilization, and a method for assisting effective diagnosis of male infertility.SOLUTION: A method for producing a selected sperm population comprises a first step of preparing a first sperm population containing, as a subpopulation, specific sperms that improve the fertilization rate of the sperm population when their content in the sperm population is reduced, and a second step of removing the specific sperms from the first sperm population to obtain a second sperm population having a reduced content of the specific sperms compared with the first sperm population.SELECTED DRAWING: None

The invention belongs to the technical field of animal breeding, and particularly relates to application of alpha-ketoglutaric acid or salt thereof in preparation of a sheep semen cryopreservation diluent. The invention discloses the application of alpha-ketoglutaric acid or salt thereof in the cryopreservation of the sheep semen for the first time, and the alpha-ketoglutaric acid or salt thereof can improve the semen quality of the frozen and unfrozen sheep semen and improve the fertilization ability of the frozen semen; such as sperm motility, sperm movement performance, sperm viability, sperm plasma membrane integrity rate, sperm acrosome integrity rate, sperm antioxidant capacity, sperm ATP level, sperm early apoptosis incidence rate, sperm in-vitro fertilized ovulation rate and / or artificial fertilization pregnancy rate, can be used for preparing the sheep sperm cryopreservation diluent.

The invention relates to a low-temperature preservation method for sheep semen, and belongs to the technical field of animal reproduction. The invention provides a method for low-temperature preservation of sheep semen, which comprises the following steps: (1) collecting breeding male sheep semen by a false vagina method to obtain the collected sheep semen; (2) mixing the sheep semen collected in the step (1) with a semen diluent and fullerene to obtain a sheep semen diluent; and (3) preserving the sheep semen diluent under the condition of 2-8 DEG C. The sheep semen is preserved according to the method, the effective application time of sperm preservation can be remarkably prolonged, the blastocyst forming rate of in-vitro fertilization embryos can be remarkably increased, and the method is applied to reproduction technologies such as artificial insemination (AI) and in-vitro fertilization (IVF) of sheep, the utilization rate of high-quality breeding rams can be increased, and genetic resource protection and efficient breeding can be promoted.

The present invention relates to transgenic ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The application discloses a one-step freezing solution for bovine in-vitro embryos and a one-step freezing method for bovine in-vitro embryos and belongs to the technical field of biotechnology.The method comprises the following steps: bovine oocyte in-vitro maturation; bovine in-vitro fertilization embryo production; after culturing the bovine in-vitro fertilization embryo for 6-8 days, the embryo is moved into freezing solution A, and after standing, the shriveled blastocyst is moved from the freezing solution A into freezing solution B, the embryo sinking to the bottom of the freezing solution B is moved into freezing solution C, is loaded into a macaroni tube, and is subjected to programmed freezing; after the freezing program is completed, the macaroni tube is put into liquid nitrogen. The bovine in-vitro embryos are frozen by using the three kinds of freezing solutions, on the premise that the conventional freezing method has low technical requirements for the operator and can be directly transplanted after thawing, the conventional freezing method is overcome, the survival rate, the hatching rate and the transplantation pregnancy rate of the embryos after thawing are low, the convenience of use and the effective integration of the high transplantation pregnancy rate are realized.

Provided herein are methods for improving the efficiency of in vitro fertilization of animals. The method comprises (a) obtaining an oocyte from the animal, wherein the oocyte comprises an oocyte body and a zona pellucida; (b) obtaining sperm from the animal; (c) maturing the oocytes in a maturing medium; (d) removing cumulus cells from the oocytes; (e) drilling in the zona pellucida of the oocyte with a laser; (f) contacting the oocyte with the sperm; (g) co-incubating the oocytes with sperms, whereby the incubation enables in vitro fertilization of the oocytes and sperms to produce animal embryos, and whereby drilling is performed in the zona pellucida of the oocytes by laser to improve the efficiency of in vitro fertilization of the production of animal embryos.

The invention provides a ricefield eel breeding technology, which relates to the technical field of ricefield eel breeding, and comprises the following steps: step 1, parent selection and anesthesia; step 2, parent spawning induction; step 3, fry incubation; step 4, temporary culture of seedlings in a constant-temperature workshop. According to the ricefield eel breeding technology, healthy male ricefield eels and healthy female ricefield eels are selected as male parents and female parents respectively, sperms are taken from the male parents, ova are taken from the female parents, in this way, defective ricefield eel sperms and ricefield eel ova are greatly reduced, and then the ricefield eel ova and ricefield eel sperms are combined in an in-vitro fertilization mode; due to the fact that the monopterus albus eggs and the monopterus albus sperms are concentrated together, fertilization of all the eggs can be completed, the fertilized eggs after fertilization are artificially cultivated and formed, the hatching rate is high, the cultivated monopterus albus fries are healthy, and the monopterus albus fries can grow healthily.

The invention relates to the technical field of animal breeding, in particular to a method for improving the in-vitro fertilization efficiency of low-concentration sperms. Comprising the following steps: treating sperms with functional proteins; the functional protein is MFG-E8; the concentration of the functional protein ranges from 250 ng / mL to 750 ng / mL. The protein MFG-E8 capable of improving the sperm motility and the in-vitro fertilization efficiency is obtained through research. In a fertilization environment, by adding the protein MFG-E8, the fertilization rate of sperms can be effectively increased, and the in-vitro fertilization efficiency under the condition of low-concentration sperms is remarkably improved. The method provided by the invention can be applied to the in-vitro fertilization technology of various mammals, and provides a new optimization strategy for the assisted reproduction technology.

The invention provides a semen adding device for in-vitro fertilization of human sperm eggs, and mainly relates to the technical field of medical equipment. A semen adding device for in-vitro fertilization of human sperms and eggs comprises an operator and a micromanipulation tube arranged in the operator, the operator comprises a supporting tube with an opening in the top face, a holding tube is in threaded connection with the opening of the supporting tube, a second through hole is formed in the center of the top face of the holding tube, and a second through hole is formed in the top face of the holding tube. A limiting assembly capable of keeping the micromanipulation tube stable is arranged in the supporting tube, a piston assembly is arranged in the holding tube, and the micromanipulation tube penetrates through the first through hole and extends out of the supporting tube. The device has the advantages that highly accurate and stable operation is achieved through cooperation of multiple components, the micromanipulation tube is doubly limited by the limiting assembly in the radial direction and the axial direction, during human sperm and egg in-vitro fertilization, sperm can be accurately added to a target position, the situation that the fertilization environment is affected due to operation errors is avoided, and the working efficiency is improved. The success rate of in-vitro fertilization is greatly improved.

A method for automated ICSI includes receiving at least one droplet containing an egg in a dish placed on a stage. The method includes using an artificial intelligence / machine learning system (AI / ML system) and an imaging system to detect a zona pellucida. The imaging system includes a microscopy system, a camera system, and a lighting system. The method includes holding the egg using a robotic microtool and lowering a robotic pipettor into the droplet. The method includes using the AI / ML system and imaging system to determine an area at which to hold the egg and positioning the robotic microtool to that area. The method includes using the AI / ML system and imaging system to instruct the robotic microtool to apply negative pressure to hold the egg to the robotic pipettor. The method includes using the AI / ML system and imaging system to determine a target location where zona ablation should be performed.

Disclosed herein are methods for determining the quality of sperm in a sperm sample. The methods may include obtaining an RNA expression profile of the sperm using PANoramic RNA Display by Overcoming RNA modification Aborted sequencing (PANDORA-seq). This disclosure also relates to methods for performing an in vitro fertilization or artificial insemination procedure and obtaining an RNA expression profile of sperm in a sperm sample.

The invention discloses an in-vitro fertilization failure risk prediction model and a construction method and a prediction method thereof, based on data of an ART treatment cycle, multi-dimensional indexes such as woman age, BMI, male seminal fluid parameters and infertility years are incorporated, logistic regression, a random forest and an XGBoost model are constructed, and performance is evaluated by adopting a nested cross validation framework (including an SMOTE technology). Results show that the logistic regression model is optimal in performance and is remarkably superior to a random forest model and an XGBoost model, key predictive factors include male age, female BMI, the total number of forward motional sperms and a sperm DNA fragmentation index, the total number of forward motional sperms and the male age are protection factors, the BMI and the sperm DNA fragmentation index are risk factors, and the risk factors are the risk factors. The C-index of the model column diagram is up to 0.722 through internal verification, and the model column diagram has the clinical distinguishing capability of medium or above. The logistic regression model is verified to have high efficiency and stability in IVF fertilization failure prediction, and can provide decision support for clinical early intervention.

The invention discloses application of salvianolic acid A in preparation of a medicine for improving the sperm function, and belongs to the technical field of biological medicine. SAA is polyphenol acid formed by caffeic acid and tanshinol, namely salvianolic acid through an ester bond and an enol bond, is a soluble component generated by many salvia species, is mainly extracted from salvia miltiorrhiza, and has the advantages that the salvianolic acid A is prepared from salvianolic acid A, salvianolic acid B and salvianolic acid C; sAA is one of the most abundant salvianolic acid in a salvia miltiorrhiza extract, a large number of studies show that SAA has a remarkable antioxidant effect, SAA can improve the function of in-vitro mature sperms, compared with a control group, after SAA is added, sperm intracellular calcium signals and hyperactivity are remarkably improved, and the occurrence rate of sperm acrosome reaction is also remarkably increased; the application of SAA can effectively improve the sperm function in the in-vitro fertilization process and improve the sperm motility of weak sperm patients, so that infertility is intervened and treated through simple non-invasive means such as artificial fertilization, and the fertility outcome of the patients is improved.

The present invention relates to transgenic ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing.

The invention relates to a yak X / Y sperm separation and enrichment method based on pH value regulation and a kit thereof, and belongs to the technical field of animal breeding and sex control. The method comprises the following steps: using an Optidyl diluent as a sperm sorting protection solution, adjusting the pH value of the Optidyl diluent through NaOH or HCl, enriching X sperms in a pH range of 6.2-6.4, and enriching Y sperms in a pH range of 6.8-7.0; the method comprises the following specific steps: diluting yak sperm, centrifugally desizing, re-suspending by using a sorting solution with corresponding pH, standing and incubating for 45-90 minutes at 35-40 DEG C and 45 DEG C, realizing natural layering according to the motility difference of X and Y sperms under different pH conditions, respectively collecting upper or lower sperms, and washing and re-suspending to obtain a sex-controllable sperm sample; the method is easy and convenient to operate, low in cost, small in sperm damage, complete in sperm structure and stable in function after separation, suitable for breeding technologies such as artificial insemination and in-vitro fertilization, capable of meeting the requirement for sexual control sperm in yak production and beneficial for optimizing the population structure and improving the breeding efficiency.

The invention provides a sheep X and Y sperm separating medium and a sorting method, and belongs to the technical field of biology. The separation liquid comprises a separation liquid, a washing liquid and requinimod R848. Wherein the washing liquid contains a protective agent, namely oroxin B, for preventing the goat sperms from being damaged by R848, so that the movement performance of the separated goat sperms can be improved, and the movement performance such as the sperm motility, the linear rate, the path rate and the linearity of the goat can be remarkably improved. In addition, the conditions of an X and Y sperm sorting system are optimized, and the X and Y sperms of the goat can be efficiently separated. The purity of X sperms sorted by the method can reach 88.1%, the proportion of female embryos after in-vitro fertilization reaches 80%, the purity of Y sperms can reach 85.7%, and the proportion of male embryos after in-vitro fertilization reaches 84%.

The present application relates to the field of animal reproductive biotechnology, and discloses a method for producing in vitro fertilized embryos by live ovum collection of snowflake beef cattle, aiming at solving the problems of uneven quality of oocytes, low efficiency of in vitro fertilization and loose connection of each technical link in the prior art. The method comprises the following steps: screening of donor cattle and superovulation treatment by decreasing injection of follicle stimulating hormone and luteinizing hormone; obtaining oocyte complex by transrectal ultrasonic guided puncture; in vitro maturation culture of the obtained oocyte; in vitro fertilization after capacitation treatment of semen by gradient centrifugation; sequential culture of the fertilized eggs to the stage of morula or blastocyst; morphological evaluation and vitrification preservation of the embryos cultured to the set stage. By the combined application of the standardized superovulation scheme, the precise ovum collection technology, the optimized maturation culture condition and the sequential culture strategy of embryos, the efficiency of embryo production can be significantly improved, and the quality of in vitro maturation of oocytes can be improved.

The present invention provides a method for improving the efficiency of in vitro fertilization in animals. The method comprises: (a) obtaining oocytes from the animal, wherein the oocytes include the oocyte body and the zona pellucida; (b) obtaining sperm from the animal; (c) maturing the oocytes in a maturation medium; (d) removing cumulus cells from the oocytes; (e) creating holes in the zona pellucida of the oocytes with a laser; (f) contacting the oocytes with the sperm; and (g) incubating the oocytes with the sperm, wherein the incubation enables in vitro fertilization of the oocytes and sperm, and an animal embryo is produced; and the efficiency of in vitro fertilization for producing an animal embryo is improved by creating holes in the zona pellucida of the oocytes with a laser. In certain embodiments, the cumulus cells are removed from the oocytes by treatment with hyaluronidase, pipetting, and / or vortexing. In one embodiment, the oocytes are placed on a microscope stage prior to step (e).

The invention belongs to the technical field of fish breeding, and particularly relates to a fish seminal plasma exosome, an extraction method and application in fish breeding. The extraction method of the fish seminal plasma exosome comprises the following steps: S1, centrifuging fish seminal fluid to obtain seminal plasma; s2, mixing the seminal plasma with PBS and BPS according to a certain volume ratio, and standing; s3, centrifuging the standing liquid to obtain a precipitate I; s4, centrifuging the precipitate I to obtain exosome precipitate; s5, adding seminal plasma into the exosome precipitate in proportion, mixing and resuspending to obtain a mixed solution I; s6, centrifuging the mixed solution I obtained in the step S5 to obtain supernate rich in exosome particles; and S7, placing the supernatant in an EPF column for centrifugation. The fish seminal plasma exosome can be used for in-vitro fertilization and artificial insemination pretreatment of fish sperms, especially in-vitro fertilization and artificial insemination pretreatment of turbot sperms, and can improve the fertilization success rate, the sperm activation efficiency and the sperm motility.