Preparation method of novel goat sperm in-vitro agonist

An agonist and goat technology, applied in the field of artificial insemination, can solve problems such as limiting the efficiency of artificial insemination of goats, and achieve the effects of easy promotion and use, easy processing and production, and improved microenvironment

Inactive Publication Date: 2019-09-17
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing studies have reported that the level of sperm motility directly affects the litter size, which limits the efficiency of artificial insemination of goats

Method used

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  • Preparation method of novel goat sperm in-vitro agonist
  • Preparation method of novel goat sperm in-vitro agonist

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A kind of preparation method of novel goat sperm in vitro agonist is characterized in that it comprises the following steps:

[0036] (1) Take adult goat testes within 8 to 10 months of age, put them into an insulated bucket of 37°C normal saline with 1% penicillin and streptomycin mixture, bring them back to the laboratory cell room on a clean bench, and place them The testicular tissue was washed three times with PBS buffer solution containing 1% penicillin-streptomycin mixture, and then put into a petri dish;

[0037] (2) Use surgical scissors to cut off the testicular albuginea, then use tweezers to peel off the inner tissue of the testis and the albuginea, remove the albuginea, retain the inner tissue of the testis, and rinse with normal saline;

[0038] (3) Cut the testis longitudinally with a scalpel to expose the testicular medulla, use tweezers to clamp 3 to 4 tissues with a diameter of 4 to 6 mm in the small septum of the testis, put them into a 6 cm petri dis...

Embodiment 2

[0047] A kind of preparation method of novel goat sperm in vitro agonist is characterized in that it comprises the following steps:

[0048] (1) Take adult goat testes within 8 to 10 months of age, put them into an insulated bucket of 37°C normal saline with 1% penicillin and streptomycin mixture, bring them back to the laboratory cell room on a clean bench, and place them The testicular tissue was washed three times with PBS buffer solution containing 1% penicillin-streptomycin mixed solution, and then put into a petri dish.

[0049] (2) Use surgical scissors to cut off the testicular albuginea, then use forceps to peel off the inner tissue of the testis and the albuginea, remove the albuginea, retain the inner tissue of the testis, and rinse with normal saline.

[0050] (3) Cut the testis longitudinally with a scalpel to expose the testicular medulla, use tweezers to clamp 3 to 4 tissues with a diameter of 4 to 6 mm in the small septum of the testis, put them into a 6 cm pet...

Embodiment 3

[0061] Embodiment 3 therapeutic effect experiment

[0062]A sheep farm in Pingdu City, Qingdao City, Shandong Province selected billy goats aged 8 to 10 months after birth, and randomly selected healthy ones to take testicular tissues, and used the agonist obtained in Examples 1 to 2 of the present invention. Randomly selected 4 breeding rams in estrus (breeding ram No. 1, breeding ram No. 2, breeding ram No. 3, and control group rams), the semen of the above 4 rams were collected by the pseudo-vaginal method, Store in a clean sterile centrifuge tube and refrigerate. After the start of the experiment, the ram semen stored in cold storage was taken out, the sperm viability was examined under a microscope and the motility rate was recorded. The semen of breeding rams No. 1 to No. 3 were added with agonist at a ratio of 1:10 (agonist: semen) and shaken well, and the control group rams were added with the same volume of normal saline and shaken well. Microscope observation and u...

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Abstract

The invention relates to a preparation method of a novel goat sperm in-vitro agonist. The method is characterized by comprising the following steps: (1) cleaning adult goat testes; (2) cutting the tunica albuginea of the testes; (3) using tweezers to clamp out tissues for washing; (4) adding a DMEM basic culture solution after digestion; (5) performing filtering and centrifuging, adding a buffer solution, and performing centrifuging; (6) adding a DMEM basic culture solution, taking cell suspension, performing centrifuging, and collecting cells; (7) adding PBS, performing centrifuging, and then adding a DMEM basic culture solution for culture; (8) carrying out cell passage, performing culturing for 8 hours, and performing storing to obtain a basic liquid I; (9) adding bill goat serum (EGS), calcium ionophore (IA), caffeine, pentoxifylline (PF), 17alpha-20beta-dihydroxyprogesterone (DHP) and heparin into a DMEM basic culture solution, and performing full and uniform mixing to obtain basic liquid II; and (10) uniformly mixing the basic liquid I with the basic liquid II according to a ratio of 5:1 to obtain the novel goat sperm in-vitro agonist. The agonist has the advantages of good application effect, low cost and easy processing and production.

Description

technical field [0001] The invention belongs to the technical field of artificial insemination, and specifically refers to a preparation method of an in vitro agonist capable of significantly improving the motility of goat sperm. The invention relates to a method for improving the motility of goat sperm, which can be used in conjunction with assisted reproductive techniques such as in vitro artificial insemination, intrauterine artificial insemination and in vitro maturation. In other aspects, the invention relates to methods for inducing maturation of goat sperm and methods of providing an improved microenvironment for goat sperm. Background technique [0002] In vitro fertilization refers to the technology in which mammalian sperm and eggs complete the fertilization process in an artificially controlled environment outside the body. It is abbreviated as IVF in English. In 1982, Brackett and others in the United States bred the world's first in vitro fertilized cattle. Sin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076
CPCC12N5/061C12N2501/91C12N2509/00C12N2500/30
Inventor 董焕声潘庆杰黄福任
Owner QINGDAO AGRI UNIV
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