55 results about "Mature sperm" patented technology

The invention relates to a factory hybrid breeding method of epinephelus fuscoguttatus and epinephelus lanceolatus, which comprises the following steps: selecting an epinephelus lanceolatus parent fish and an epinephelus fuscoguttatus parent fish for parent fish cultivation, artificially adjusting and controlling the cultivation water temperature of the epinephelus lanceolatus parent fish from 23 DEG C to 20 DEG and from 20 DEG to 30 DEG, controlling the cultivation water temperature of the epinephelus fuscoguttatus parent fish from 23 DEG C to 20 DEG and from 20 DEG to 28 DEG and taking mature sperm of the epinephelus lanceolatus parent fish in May-June of a next year for standby; and choosing a female fish of which an abdomen obviously expands, and an ovary develops to the stage IV, injecting a parturifacient, fishing an epinephelus fuscoguttatus female fish for artificial egg stripping after 36-hour effector phase, then mixing an collected epinephelus fuscoguttatus fish egg with the collected epinephelus lanceolatus mature sperm according to the volume ratio of 1000:1, adding sea water to finish fertilization, putting a fertilized egg into an incubation tank, collecting a floating eyed egg after about 20 hours and sterilizing by adopting ozone with a concentration of 0.3-0.5 ppm. Through the factory hybrid breeding method, the hybrid breeding of the epinephelus fuscoguttatus and the epinephelus lanceolatus is realized.

The invention discloses a coated carrier and the application thereof in the method for detecting maturity of sperms and separating mature sperms noninvasively; the coated carrier is a carrier coated by hyaluronic acid, or hyalurate or hyaluronic acid derivatives; the detecting method is as follows: preparing specimens; loading the specimens to the coated carrier and covering with microscopic glass; reacting for 5-60 minutes at room temperature; observing by a microscope and counting; and calculating the combination ratio of hyaluronic acid of active sperms. The separating method is as follows: preparing specimens; loading the specimens to the coated carrier, reacting for 5-120 minutes at the temperature of 18-37 DEG C; and collecting the combined sperms. The coated carrier has the advantages of precise and reliable results, simple and fast operation, low cost and good safety performance, and can be used for estimating sperms quality, sperms maturity and analysis of the causes of sterility disease of mails, and is applicable to assisting reproduction or reproduction research center to safely collect mature and active sperms, so that the goal of improving embryo quality greatly is achieved.

A method of obtaining normal mature sperms of eel relates to a method of obtaining normal mature sperms of eel requiring manual adjusting and controlling to obtain normal mature sperms of eel. The invention comprises: performing backgrounding domestication and food calling on the received catadromous eel parent, causing wild eel to adapting manual breeding environment, increasing corporeity of the eel parent through food calling, storing more matter and energy, preparing for entering the seawater, characterized in that, the eel parent is performed with salinity domestication after being bred for 15 days under manual breeding environment, salinity domestication is transferred from low concentration to sea water; water temperature of a culture pond for the eels is controlled between 18 to 25 DEG C, light is shielded in the day and moonlight is seen at night, flowing water stimulation is performed at night, oxygen concentration in the water keeps more than 6 mg / l, in weak alkalinity water, content of ammonia and nitrogen is not more than 0.02 mg / l and 0.05 mg / l, eel males discharge seminal fluid naturally, normal mature sperms are obtained. The invention is used for fully artificial propagation for the eels.

The invention belongs to field of fish breeding and aquaculture, which in detail relates to a method of artificial crossing with summer-bothus and tooth-Bothus and breeding. It comprises following steps: (1) taking summer-bothus as male parent and tooth-bothus as female parent, reinforcing the nutrient and environmental condition to promote simultaneous gonadal maturity, getting mature sperm and ovum; (2) crossing fertilized ovum and hatching; (3) culturing the hybrid body and young fish; (4) providing systematic bait in culturing period. The invention not only provides material for deseription improvement for bothus and soles, but also the lies foundation of adding a new breeding species for sea fish cultivating.

The invention provides a method for breeding tridentiger trigonocephalus by artificial induced spawning and insemination, which relates to the artificial propagation of gobies. The method required tobe provided for artificially breeding the tridentiger trigonocephalus comprises the steps of ripe fish screening, maturity acceleration, insemination, incubation and larval fish culture, wherein the maturity acceleration of ripe fishes uses luteotropin to release hormone analogues, domperidone maleate and chorionic gonadotropin, and is characterized in that the three hormones are used twice for the maturity acceleration; in the maturity acceleration, the water temperature of culture is between 18 and 21 DEG C, the salinity is between 2 and 6, the dissolved oxygen is between 3.5 and 6.5mg / L, and the pH is between 7.5 and 9.0; and after the second time of the maturity acceleration, in 8 to 10 hours, mature ova and mature sperms are selected for artificial insemination which comprises the following steps that: the activated sperms are poured on the ova for sufficient contact so as to obtain fertilized ova; the fertilized ova are placed in a hatchery pond for incubation; and the hatched larval fish is placed in a culture pond for incubation, and chlorella and yolk powder of eggs are placed in water from the third day after the larval fish is hatched to feed food for the larval fish. The method is used for the artificial breeding of the tridentiger trigonocephalus.

The invention relates to an artificial fertilization and incubation method of sinocyclocheilus grahami, which belongs to the technical field of fishery product culture. The method comprises the steps of dry fertilization and artificial incubation. Intramuscular injection is performed on sexually matured parent sinocyclocheilus grahami with DOM with the use level of 1 mg for per kilogram body weight and luteinizing hormone releasing hormone A2 with the use level of 1 mug for per kilogram body weight; 24 hours later, the mature germ cell and the mature sperm are slightly squeezed out from the abdomens of female sinocyclocheilus grahami and male sinocyclocheilus grahami respectively; the germ cell and the sperm are slightly stirred for 30 seconds in a vessel by chicken feather; clear water is added into the germ cell and the sperm and stirring is performed for 20 seconds; the germ cell and the sperm are cleaned for three times by clear water; the germ cell and the sperm are uniformly spilled onto a palm sheet which is cleaned and sterilized and put into 5 ppm mould solution to be dipped and sterilized for 15 munities, then the germ cell and the sperm are taken out and put into a 20 m or 50 incubating pond to be incubated; an oxygen pump is used for oxygenation to performing incubation under the condition of 18 to 20 DEG C water temperature and 7.5 to 8.0 pH, and after 150 hours, the incubation is accomplished. The method has the advantages that the cost is low, the operation is simple, the method can be put into industrial and intensive production, and the fertilization rate and the incubation rate are high.

The invention provides an animal model with male reproductive disorders of a non-human mammalian animal, as well as a preparation method and application of the animal model, and further identifies the functions of a Prss37 gene and a protein. The Prss37 gene of the animal model is inactivated, thereby affecting the appearance of ADAM3 in mature sperms produced by the animal model, indirectly affecting the combination of the sperms-zona pellucida and the migration of the sperms from the uterus to the oviduct and resulting in the serious male reproductive disorders. The model, the Prss37 gene and the protein thereof can be used for screening ADAM3 ripening accelerators, as well as sterility infertility treatment agents or contraceptive medicaments.

The invention discloses a method for complete inactivating germ genetic substance for yellow croaker, which can be used for research and production foryellow croaker gynogenesis and whole femal seed induction. The method comprises following steps: diluting the mature sperm of yellow croaker for 2- 100 times with precooled nutriment- containing diluting liquid; putting it on bottom culturing utensil; irradiating with violet light and impregnating with spawn and incubating; then identifying the effect of sperm genetic inactivation. The invention is characterized by simple method, convenient operation, and 100% of the inactivating rate for sperm genetic substance.

The present invention provides methods, devices, and kits for separating and selecting top sperm from a sperm sample of a subject. In one aspect, for example, such a method can include removing a portion of negatively charged protein from sperm in the sperm sample, immobilizing the sperm, electrophoretically separating the sperm, and selecting mature sperm based on electromotility properties.

A flow cytometry method and kit is provided for quantifying an absolute number of mature sperm cells in a semen sample, the method comprising (a) determining a total particle number (T); (b) determining a number of DNA comprising particles (D); (c) determining a number of cells which are not mature sperm cells (B) and; (d) determining a background signal (A); wherein steps (a) to (d) are not effected in a single aliquot of the semen sample, and whereas the number of mature sperm cells in the semen sample is equal to [(T×D) / 100]−(A+B), thereby quantifying the absolute number of mature sperm cells in the semen sample.

A flow cytometry method and kit is provided for quantifying an absolute number of mature sperm cells in a semen sample, the method comprising (a) determining a total particle number (T); (b) determining a number of DNA comprising particles (D); (c) determining a number of cells which are not mature sperm cells (B) and; (d) determining a background signal (A); wherein steps (a) to (d) are not effected in a single aliquot of the semen sample, and whereas the number of mature sperm cells in the semen sample is equal to [(T×D) / 100]−(A+B), thereby quantifying the absolute number of mature sperm cells in the semen sample.

Provided are methods of in vitro maturation of human spermatogonium, comprising culturing the spermatogonium in a three-dimensional methylcellose culture system (MCS) under conditions capable of differentiating said human spermatogonium into an elongated spermatid, thereby in vitro maturing the human spermatogonium. Also provided is an in vitro matured sperm obtainable according to the method of the invention and methods of treating subjects in need of mature sperm cells.

The invention belongs to the field of aquaculture, and particularly relates to an artificial control method for inducing the synchronous maturing of the gonads of male paralichthys dentatus parent and female paralichthys olivaceus parent. In the method, the male paralichthys dentatus parent is subjected to full-illumination environmental control in autumn and winter to suppress the gonad development of the male paralichthys dentatus parent; and in the spring of the next year, the full-illumination environment is changed into short-illumination environment, and the male paralichthys dentatus parent and the female paralichthys olivaceus parent are subjected to gonad development control at the same time to realize the gonad maturing of the male paralichthys dentatus parent. The method provided by the invention can obtain mature sperms and ova at the same time, prolongs the germ production period of the male paralichthys dentatus parent, increases the sperm quantity, and lays a technological base for obtaining batch high-quality sperms and ova in the large-scale offspring seed production of hybrid flounder.

The invention discloses an artificial insemination method for Filicinae, and belongs to the technical field of plant artificial insemination. In order to solve the problem of low seedling rate of pteridophytes, the invention provides an artificial insemination method of Filicinae, which comprises the following steps: inoculating spores into a culture medium, obtaining female gametophytes and malegametophytes of the Filicinae by using a conventional spore culture method, after a sexual organ is mature, sucking mature sperms and a prepared induction liquid, sequentially dropwise adding the mature sperms and the prepared induction liquid to a mature archegonium of the female gametophyte, and enabling the sperms to enter the archegonium to be combined with ova under the guidance of the induction liquid, so as to complete artificial insemination. According to the sperm taking and artificial directional insemination technology provided by the invention, the fertilization rate of the gametophyte can reach 30%-50% and is 3-5 times that under conventional culture conditions, the phenomenon that a single gametophyte has two embryonic developments often occurs, and the hybridization technology of pteridophytes is simpler, more convenient and easier to implement.

Methods, systems and apparatus for preparing sperm for ICSI by combining selective enrichment benefits of sperm gradient preparation, sperm swim-up preparation and hyaluronan-binding for selection of mature sperm into a single processing unit. The single processing unit of the invention reduces operator labor steps and time, is efficient, easy to use, offers quick processing time and provides highly motile, mature and specific individual sperm that retain desired sperm characteristics selected by each individual selection principal that are ready for capture and use in ICSI.

The invention discloses a cytology division method of a scophthalmus maximus spermiogenesis phase. The method comprises the following steps: fixing, pre-dyeing, dehydrating and embedding, dyeing by uranyl acetate, dyeing by lead citrate, and slicing preparing and analysis. A scophthalmus maximus spermiogenesis process is divided into 8 phases: comprising sperm cell I-VII stage and mature sperm. The method has the beneficial effects that the cytology division method overcomes the problem that during common histological slice observation, sperm cells at different metamorphosis periods cannot bedistinguished due to limited magnification times, pre-dyeing is helpful for increasing dyeing uniformity, tissue structure definition, and tissue dyeing contrast grade, dyeing effect can be optimized,and image definition of the tissue structure of the sperm cells can be obviously enhanced; the cytology division method can fill the research blank of phase division for the flounder paralichthys spermiogenesis, and can be used for cytology division of the spermiogenesis phase of the other seawater fish.

The invention discloses a device for selecting sperms coated with hyaluronic acid and hyaluronates or hyaluronic acid derivatives. The device comprises a small groove (21) integrally formed in the bottom in a petri dish body (2); the small groove (21) is coated with hyaluronic acid, and hyaluronates or hyaluronic acid derivatives capable of being combined with mature sperms, and is higher than the coated hyaluronates or hyaluronic acid derivatives; a plurality of small lugs (22) which allow the petri dish body (2) to be taken conveniently and avoid contact of a petri dish opening of the petri dish body (2) are distributed on the outer side of the petri dish body (2); a convex step (23) for preventing the outer bottom of the petri dish body (2) from being worn is arranged at the outer bottom of the petri dish body (2); an angle formed between the highest part of the edge of the petri dish body (2) and the bottom of the small groove (21) is 5-23 degrees. The invention further discloses an application method of the device. The device is easy to use and convenient to operate; through the adoption of the device, the mature sperms can be selected conveniently without destroying morphology, structure and vitality of the sperms.

The present invention provides methods, devices, and kits for separating and selecting top sperm from a sperm sample of a subject. In one aspect, for example, such a method can include removing a portion of negatively charged protein from sperm in the sperm sample, immobilizing the sperm, electrophoretically separating the sperm, and selecting mature sperm based on electromotility properties.

The invention belongs to biotechnology and medical field and discloses a new human epididymis specific expression sperm binding protein HEL-172 and preparation and application thereof. The invention discloses an amino acid sequence of the HEL-172 protein and provides a recombination expression vector carrying a DNA sequence for encoding the protein. The invention further discloses a method for preparing the protein and detecting the biological function of the HEL-172 protein. The protein is positioned at the back region of the head of a mature sperm, has expression in epididymis and spermary, heart, liver, spleen, kidney and stomach. A band of about 34 KD is detected in the epididymis by using Western blot. The HEL-172 protein can be used as a target protein for the research and development of immunity acyeterion as well as clinical diagnosis and treatment of male infertility. The relevant gene or protein detecting method based on the protein HEL-172 can be widely applied to the reproductive medicine research field.

A method of obtaining normal mature sperms of eel relates to a method of obtaining normal mature sperms of eel requiring manual adjusting and controlling to obtain normal mature sperms of eel. The invention comprises: performing backgrounding domestication and food calling on the received catadromous eel parent, causing wild eel to adapting manual breeding environment, increasing corporeity of the eel parent through food calling, storing more matter and energy, preparing for entering the seawater, characterized in that, the eel parent is performed with salinity domestication after being bred for 15 days under manual breeding environment, salinity domestication is transferred from low concentration to sea water; water temperature of a culture pond for the eels is controlled between 18 to 25 DEG C, light is shielded in the day and moonlight is seen at night, flowing water stimulation is performed at night, oxygen concentration in the water keeps more than 6 mg / l, in weak alkalinity water, content of ammonia and nitrogen is not more than 0.02 mg / l and 0.05 mg / l, eel males discharge seminal fluid naturally, normal mature sperms are obtained. The invention is used for fully artificialpropagation for the eels.

The invention belongs to the biotechnology and the medical field and discloses a new human epididymal specific expression sperm binding protein HEL-28 as well as preparation and application thereof. The invention discloses an amino acid sequence of the HEL-28 protein and provides a recombinant expression vector which carries a DNA sequence used for encoding the protein. The protein is located in the back region of the head of a mature sperm, and an RT-PCR result indicates that the protein has low expression level in an epididymis and various tissues of a humna body. A band of 187 KD is detected in the body part of the epididymis by using Western blot. The HEL-28 protein can be used as a target protein for developing an immunological contraceptive and clinically diagnosing and treating male infertility. A method for detecting a corresponding gene or protein based on development of the HEL-28 protein can be widely used for the research field of reproductive medicine.

The invention belongs to the biotechnical field, and relates to an immunogen for preparing an LRRC52 polyclonal antibody. The immunogen is a polypeptide, and the amino acid sequence of the immunogen is CQNNRIREVMDYTFI. The LRRC52 polyclonal antibody prepared by using the immunogen has the advantages of high titer and good specificity. The antibody can substantially inhibit the generation of sperm potassium current after incubated with mouse or human sperms in order to substantially influence the physiologic functions of mouse and human mature sperms. The antibody can be used for researching the physiologic adjusting mechanism of the sperms as a tool medicine, and can also be used for developing novel female contraceptives as a candidate molecule.

The invention belongs to the field of biotechnology and medicine, and discloses a new human epididymis specific expression sperm binding protein HEL-72 as well as preparation and application thereof. The invention discloses an amino acid sequence of the HEL-72 protein and provides a recombination expression carrier carrying the DNA sequence for encoding the protein. The protein is located in the primary segment and tail segment at the tail part of a mature sperm. A RT-PCR result shows that the gene is lowly expressed in the epididymis, highly expressed in the tissues of testis, kidney and stomach, and is negative in the heart and liver. A band of about 30 KD is detected in the corpus epididymidis part with Western blot. The HEL-72 protein can be used as a target protein for researching and developing immunity contraceptive as well as clinical diagnosis and treatment for male sterility. A method for detecting a corresponding gene or protein based on development of the HEL-72 protein can be widely used for the research field of reproductive medicine.

The invention belongs to the field of biotechnology and medicine, and discloses a new human epididymis specific expression sperm binding protein HEL-97 as well as preparation and application thereof. The invention discloses an amino acid sequence of the HEL-97 protein and provides a recombination expression carrier carrying the DNA sequence for encoding the protein. The protein is located in the middle rear and tail parts of the head of a mature sperm. A RT-PCR result shows that the gene is weakly or rarely expressed in the head and tail part of the epididymis and other tissues of testis, heart, liver, spleen, kidney and stomach. A band of about 28 KD is detected in the corpus epididymidis part with Western blot. The HEL-97 protein can be used as a target protein for researching and developing immunity contraceptive as well as clinical diagnosis and treatment for male sterility. A method for detecting a corresponding gene or protein based on development of the HEL-97 protein can be widely used for the research field of reproductive medicine.

The invention provides a mountainous area loach parent cultivation and artificial induce spawning method, and an incubation system. The mountainous area loach parent cultivation and artificial induce spawning method includes the following steps: selecting loach parents, and breeding the female loach parents and male loach parents in different pools; performing long water flow stimulation to enhance gonad development one month before a mating season; dissolving 400-500 IU / kg fish chorionic gonadotrophin HCG and 6-8 ug / kg luteinizing hormone releasing hormone LHRH_A2 in 4-5 mg / kg domperidone DOM to be used as an inducing agent; collecting mature sperms and eggs in a period after induce spawning; and using a dry method and a semi-dry method to perform artificial insemination. The problem of the quality of loach parent cultivation in mountainous area pool resource characteristics can be solved; a mountainous pool loach artificial cultivation and high efficiency reproduction flow is formed so as to perfect the prior art, and the breeding benefits can be increased.

The invention belongs to the biotechnical field, and relates to an immunogen for preparing an LRRC52 polyclonal antibody. The immunogen is a polypeptide, and the amino acid sequence of the immunogen is CQNNRIREVMDYTFI. The LRRC52 polyclonal antibody prepared by using the immunogen has the advantages of high titer and good specificity. The antibody can substantially inhibit the generation of sperm potassium current after incubated with mouse or human sperms in order to substantially influence the physiologic functions of mouse and human mature sperms. The antibody can be used for researching the physiologic adjusting mechanism of the sperms as a tool medicine, and can also be used for developing novel female contraceptives as a candidate molecule.

The invention belongs to the biotechnology and the medical field and discloses a new human epididymal specific expression sperm binding protein HEL-47 as well as preparation and application thereof. The invention discloses an amino acid sequence of the HEL-47 protein and provides a recombinant expression vector which carries a DNA sequence used for encoding the protein. The protein is located in the neck region of a mature sperm, and an RT-PCR result indicates that the protein mainly has expression in an epididymis, a testis, a liver and tissues. A band of 47 KD is detected in the epididymis by using Western blot. The HEL-47 protein can be used as a target protein for developing an immunological contraceptive and clinically diagnosing and treating male infertility. A method for detecting a corresponding gene or protein based on development of the HEL-47 protein can be widely used for the research field of reproductive medicine.

The invention belongs to the field of biotechnology and medicine, and discloses a new human epididymis specific expression sperm binding protein HEL-61 as well as preparation and application thereof. The invention discloses an amino acid sequence of the HEL-61 protein and provides a recombination expression vector carrying the DNA sequence for encoding the protein. The protein is located in the middle tail area of the head of a mature sperm. A RT-PCR result shows that the gene is mainly expressed in the tissues of epididymis, testis, liver and spleen, and is negative in the kidney and stomach expression. A band of about 21 KD is detected in the corpus epididymidis body part with Western blot. The HEL-61 protein can be used as a target protein for researching and developing immunity contraceptive as well as clinical diagnosis and treatment for male sterility. A method for detecting a corresponding gene or protein based on development of the HEL-61 protein can be widely used for the research field of reproductive medicine.