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Glutathione-contained boar semen cryopreservation liquid and cryopreservation method thereof

A technology of cryopreservation solution and glutathione, which is applied in the field of livestock breeding, can solve the problems of low survival rate of frozen semen, loss of lipid substances, low conception rate, etc., and achieve low survival rate, low economic cost, and easy promotion Applied effect

Inactive Publication Date: 2010-01-13
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, pig frozen semen has been difficult to be widely used in production. There are two reasons. One is that the ratio of cholesterol to phospholipid in the plasma membrane of pig sperm cells is significantly lower than that of herbivores, and the ion permeability of the sperm membrane is enhanced during the freezing process. The loss of lipids leads to low survival rate of frozen semen and low conception rate
Second, pig sperm are often damaged by free radicals and oxidative stress during freezing, which affects the survival rate and fertilization ability of pig sperm after freezing
But so far, there has been no report about the use of glutathione in the field of livestock reproduction technology, especially for the cryopreservation of pig semen

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of pig semen cryopreservation solution and freezing dilution containing glutathione

[0045] Preparation of pig semen cryopreservation solution containing glutathione:

[0046]Accurately weigh 400mg of glutathione, first use 50mL of BTS diluent (each 1000mL is prepared according to the following formula: glucose 37g, sodium citrate 6g, potassium chloride 0.75g, sodium bicarbonate 1.25g, ethylenediaminetetraacetic acid 1.25g g, 100000 units of penicillin, 100 mg of streptomycin, supplemented with deionized water to 1000 mL) to dissolve, then add 8 g of lactose, 20 mL of egg yolk (obtained from eggs), 8 mL of glycerin and 0.5 g of lauryl sulfate Sodium, then add BTS diluent to 100mL, stir and mix again. After filtration and disinfection with a 0.22 μM microfiltration membrane, store at 5°C for use.

[0047] Preparation of frozen dilutions of porcine semen containing glutathione:

[0048] Accurately weigh 400 mg of glutathione, first dissolve it w...

Embodiment 2

[0049] Embodiment 2: Pig semen cryopreservation liquid containing glutathione is used for large gram pig semen cryopreservation

[0050] Large gram boars with high fecundity and strong physique are selected, and semen is collected by hand holding method. Collect the sperm-rich part, filter through 4 layers of sterilized gauze to remove the colloid, measure the semen volume immediately, and check the sperm motility rate, collect about 50 mL of semen with a sperm motility rate above 75%, and dilute it twice with isothermal BTS diluent. Allow to stand at 15°C for 2 hours, and then centrifuge at 1000 rpm for 10 minutes.

[0051] After the semen is centrifuged, the semen is removed, and the sperm precipitation is diluted with a glutathione-containing porcine semen freezing diluent (Example 1), which is a cryopreservation solution of the present invention without glycerin and sodium lauryl sulfate, because glycerin Low temperature conditions have toxic and side effects on sperm. Af...

Embodiment 3

[0052] Example 3: Thawing of large gram pig frozen semen

[0053] In order to detect the implementation effect of the patent of the present invention, after the large gram pig semen was processed in turn according to the method of Example 1 and Example 2, the frozen tubes of the test group and the control group were respectively placed in a 37°C water bath to thaw for 10 seconds , the viability rate was detected under the microscope, among which the viability rate of the frozen semen in the control group was 30.2%, and the sperm viability rate in the test group was 36.7%, which was 6.5% higher than that in the control group. Statistical analysis showed that there was a difference between the two significantly. The results confirmed that the cryopreservation of large gram pig semen with glutathione-containing pig semen cryopreservation solution can effectively improve the survival rate after thawing.

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PUM

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Abstract

The invention provides glutathione-contained boar semen cryopreservation liquid and a cryopreservation method thereof. The boar semen cryopreservation liquid uses diluent which is generally suitable for boar semen cryopreservation as a substrate and also comprises 2-6 mg / ml of glutathione, 5-11 mg / ml of lactose, 10-30 percent (v / v) of yelk, 5-10 percent (v / v) of glycerin and 0.5-2 mg / ml of sodium lauryl sulfate. The invention applies the glutathione to the boar semen cryopreservation, can effectively remove redundant free radicals generated in a refrigeration process, solves the problem of low survival rate of the prior boar semen cryopreservation, has lower economic cost and is convenient to popularize and apply in production.

Description

(1) Technical field [0001] The invention relates to a pig semen cryopreservation solution containing glutathione and a cryopreservation method thereof, belonging to the technical field of livestock reproduction. (2) Background technology [0002] In my country, pigs are the most important livestock, and pork production accounts for about 70% of the total meat in the country. In recent years, with the improvement of people's living standards, the direction of pig breeding has changed from mainly focusing on the improvement of growth rate and lean meat percentage to improving the quality of pork and the flavor of meat. The traditional breeding method has a long generation interval and cannot meet the market demand. However, the pig frozen semen technology can make full use of excellent breeding stock, shorten the generation interval of fine breed selection, and quickly establish a new hybrid of breeding pigs and commercial pigs that meets market requirements. breeding system....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 王争光俞颂东陈阿琴董文艳
Owner ZHEJIANG UNIV
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