A method for improving the effect of in vitro fertilization with thawed pig sperm without treating infertility

An in vitro fertilization, pig sperm technology, applied in biochemical equipment and methods, preservation of human or animal body, embryo cells, etc., can solve the problems of high polysperm fertilization rate, poor embryo quality, poor oocyte maturation quality , to achieve the effect of improving fertilization effect and enhancing activity

Active Publication Date: 2022-01-11
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poor oocyte maturation quality, high polysperm fertilization rate and poor embryo quality are the main problems faced by pig IVP

Method used

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  • A method for improving the effect of in vitro fertilization with thawed pig sperm without treating infertility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for improving the effect of in vitro fertilization of pig sperm after thawing without treating infertility, comprising the following steps:

[0026] (1) The frozen semen is taken out and then thawed, wherein the frozen base fluid used for semen freezing is made of the following components by weight: 200 parts of glucose, 300 parts of lactose, 10 parts of sodium bicarbonate, 5 parts of penicillin sodium, 8 parts of streptomycin sulfate, 1800 parts of egg yolk, 7000 parts of three-distilled water, wherein phenethyl caffeate and polyvinylpyrrolidone are added to the frozen base liquid, and the molar concentration of caffeic acid phenethyl in the frozen base liquid is 30 μmol / L, the mass fraction of polyvinylpyrrolidone in the freezing base liquid is 0.4%;

[0027] (2) Wash the thawed semen with fertilization operation solution and centrifuge twice, then wash and centrifuge the centrifuged pellet once with mTBM capacitation solution, and add mTBM capacitation solu...

Embodiment 2

[0033] A method for improving the effect of in vitro fertilization of pig sperm after thawing without treating infertility, comprising the following steps:

[0034] (1) The frozen semen is taken out and then thawed, wherein the frozen base liquid used for semen freezing is made of the following components by weight: 250 parts of glucose, 350 parts of lactose, 13 parts of sodium bicarbonate, 6 parts of penicillin sodium, 10 parts of streptomycin sulfate, 2000 parts of egg yolk, 8000 parts of triple distilled water, wherein caffeic acid phenethyl and polyvinylpyrrolidone are added to the freezing base liquid, wherein the molar concentration of caffeic acid phenethyl in the freezing base liquid is 40 μmol / L, the mass fraction of polyvinylpyrrolidone in the freezing base liquid is 0.5%;

[0035] (2) Wash the thawed semen with the fertilization solution and centrifuge twice, then wash and centrifuge the centrifuged pellet once with mTBM capacitation solution, add mTBM capacitation...

Embodiment 3

[0041] A method for improving the effect of in vitro fertilization of pig sperm after thawing without treating infertility, comprising the following steps:

[0042] (1) The frozen semen is taken out and then thawed, wherein the frozen base liquid used for semen freezing is made of the following components by weight: 300 parts of glucose, 400 parts of lactose, 15 parts of sodium bicarbonate, 7 parts of penicillin sodium, 12 parts of streptomycin sulfate, 2200 parts of egg yolk, 9000 parts of triple distilled water, wherein caffeic acid phenethyl and polyvinylpyrrolidone are added to the freezing base liquid, wherein the molar concentration of caffeic acid phenethyl in the freezing base liquid is 50 μmol / L, the mass fraction of polyvinylpyrrolidone in the freezing base liquid is 0.6%;

[0043] (2) Wash the thawed semen with the fertilization solution and centrifuge twice, then wash and centrifuge the centrifuged pellet once with mTBM capacitation solution, add mTBM capacitation...

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Abstract

The invention discloses a method for improving the in vitro fertilization effect of thawed swine sperm without treating infertility, comprising the following operation steps: (1) taking out the frozen semen and then thawing it; (2) using the thawed semen with The fertilization operation solution was washed and centrifuged twice, and then the centrifuged sediment was washed and centrifuged once with mTBM capacitative solution, and then resuspended and cultured to obtain capacitative semen; (3) The ovary from which the follicular fluid was extracted was used to pick out the cumulus-egg under a microscope Mother cell complex, take out the oocyte after culturing, and remove the cumulus; (4) add capacitative semen to each fertilization droplet, put it in the incubator and wash off the sperm attached to the oocyte, and in vitro fertilization Cultivation is complete. In the present invention, the corresponding concentration of caffeic acid phenethyl ester and polyvinylpyrrolidone are combined in the freezing base solution I, and the synergistic effect of the two can effectively improve the vitality, plasma membrane integrity rate and DNA integrity rate of thawed pig sperm. Thus, the in vitro fertilization effect of thawed porcine sperm is improved.

Description

technical field [0001] The invention relates to the technical field of cryopreservation of pig sperm, in particular to a method for improving the in vitro fertilization effect of thawed pig sperm without treating infertility. Background technique [0002] The cryopreservation of semen can effectively preserve the genetic breeding and germplasm resources of organisms [8]. With the rapid development of pig frozen semen production technology and artificial insemination technology, the conception rate and number of piglets of breeding pigs bred with frozen semen have been effectively improved, which is close to the breeding level of fresh semen. However, due to many reasons such as the difficulty in the production of pig frozen semen, the low survival rate after freezing and thawing, and the low conception rate, the research on pig semen freezing technology is still not fully mature. In general, there are still many problems to be solved urgently to perfect the preparation tech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073C12N5/076A01N1/02
CPCC12N5/061C12N5/0604A01N1/0226A01N1/0221A01N1/0215A01N1/021C12N2500/14C12N2500/12C12N2500/34C12N2500/30C12N2500/32C12N2500/60C12N2501/998
Inventor 钱芙蓉李兆华张志彬于永生高一菅芯蕊王珺曹阳张立春昭日格图孙华王俊恒李彦春刘向光金庆国柳海星高见红马腾飞孟庆祝丁德李闯张鑫陆永辉祝永凯
Owner JILIN ACAD OF AGRI SCI
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