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Sheep X/Y sperm separation freezing sperm as well as preparation method and use thereof

A technology for freezing semen and production methods, which is applied in the directions of biochemical equipment and methods, applications, microorganisms, etc., to achieve the effects of improving separation efficiency, high vitality, and improving breeding effect.

Active Publication Date: 2009-01-21
内蒙古赛科星繁育生物技术(集团)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when using this method to separate sheep X / Y sperm, there is still a lot of room for improvement in terms of separation speed, separation accuracy, buffer use, and sperm viability after cryopreservation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Production of sheep X frozen semen and artificial insemination to control lambing sex:

[0036] 1) Collect 4 mL of fresh semen with a total sperm count of 4 billion from a healthy Mongolian ram, and add antibiotics (Tylosin: 0.103 mg / mL, Gentamicin: 0.508 mg / mL, Linco-spectromycin Linco-spectin: 0.6mg / mL), incubate at 18-20°C, use a sperm density-motility analyzer to detect the concentration and motility of fresh semen, after testing, the sperm of the semen 85% motility, transported back to the lab for 5 hours for X / Y sperm separation.

[0037] 2) According to the converted volume of semen concentration, take 4×200 million sperm (for 2 sperm separators) and centrifuge to remove the seminal plasma, add 10 μg / mL Hochest 33342 and stain in a warm water bath at 24°C for 20 minutes.

[0038] 3) The DNA content difference of sheep X / Y sperm separation is set to 3.2-3.4%, and the sperm flow cytometry separator (SX-MoFlo flow cytometry machine of American XY company) is set ac...

Embodiment 2

[0056] Production of goat X frozen semen and artificial insemination, embryo transfer to control lamb sex:

[0057] 1) A total of 6 mL of fresh semen with a total sperm count of 4.8 billion was collected from two healthy cashmere goat rams, and an appropriate amount of antibiotics (Tylosin: 0.103 mg / mL, gentamicin Gentamicin: 0.508mg / mL, Linco-spectromycin: 0.6mg / mL), keep warm at 18-20°C. The concentration and motility of the fresh semen were tested with a sperm density-motility meter. After testing, the motility of the semen was 83%, and it was transported back to the laboratory for 6 hours of X / Y sperm separation.

[0058] 2) According to the converted volume of semen concentration, take 4×200 million sperm (for 2 sperm separators) and centrifuge to remove the seminal plasma, add 10 μg / mL Hochest 33342 and stain in a warm water bath at 24°C for 20 minutes.

[0059] 3) The DNA content difference of sheep X / Y sperm separation is set at 3.2-3.4%, and the parameters of the spe...

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PUM

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Abstract

The invention relates to sheep X / Y sperm-separated frozen semen, the production method and application thereof. The sheep X / Y sperm-separated frozen semen provided by the invention contains a frozen balance solution A, a frozen balance solution B including 12 percent glycerin cryoprotectant and a sheep X or Y sperm-separated frozen semen. The invention applies the difference of DNA content of sheep X and Y sperms and separates X and Y sperms through flow cytometry so as to manufacture the sex-control frozen semen and the frozen storage condition is 20 to 30min treatment under -90 to 120 DEG C. The frozen semen provided by the invention controls the lamb sex through artificial insemination or embryo transplantation. With high separation efficiency and sex control accuracy up to 90 percent, the frozen semen is suitable for industrialization production and culture, thus greatly increasing the culturing effect.

Description

technical field [0001] The invention relates to a livestock X / Y sperm separation technology, in particular to sheep X / Y sperm separation frozen semen and its production method and application. Background technique [0002] Livestock-based frozen semen, artificial insemination and in vitro fertilization techniques are widely used in production breeding and reproductive biology research. Under normal circumstances, the progeny of sheep (sheep and goat) breed male and female respectively account for 50%, but in actual production, in order to improve the breeding effect of sheep (sheep and goat), more breeding ewes are needed for specific strains ( Sheep, goat breeding and group expansion) or male lambs (commercial mutton production, commercial cashmere production), so it is necessary to control the sex of offspring of sheep (sheep and goats) by separating X / Y sperm. In addition, separated sperm usually need to be cryopreserved before insemination, and different species have di...

Claims

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Application Information

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IPC IPC(8): A61K35/52C12N5/06A61P15/00A01N1/02
Inventor 李喜和王建国钱松晋胡树香周文忠
Owner 内蒙古赛科星繁育生物技术(集团)股份有限公司
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