Hu sheep semen cryopreservation liquid containing exosome and cryopreservation method

A cryopreservation, exosome technology, applied in the preservation of human or animal bodies, cell dissociation methods, biochemical equipment and methods, etc., can solve problems such as difficulty in meeting artificial insemination, few effective sperm, and poor quality , to achieve the effect of satisfying artificial insemination, reducing damage and improving insemination ability

Inactive Publication Date: 2022-03-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing sheep frozen semen has a low viability after thawing and there are many "aging" sperm and weak dead sperm. The effective sperm is less, the vitality is low, and the quality is poor. It is difficult to meet the needs of artificial insemination.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] In this embodiment, a Hu sheep semen cryopreservation solution contains the following components: Tris 30g / L, citric acid 16g / L, fructose 8g / L, sorbitol 10g / L, glycerol 7.5v / v%, BSA 3g / L, mesenchymal stem cell freeze-dried powder 0.3g / L, penicillin 1 million IU / L, streptomycin 1g / L.

[0028] The preparation method of the Hu sheep mesenchymal stem cell freeze-dried powder is as follows:

[0029] (1) Collect Hu sheep umbilical cord under aseptic conditions;

[0030] (2) Wash the residual blood, separate and obtain Wharton's jelly, then cut it into small pieces, add type II collagenase at a concentration of 1 mg / mL, digest at 37°C for 6 hours to obtain a single cell suspension, and add PBS to stop digestion , centrifuge, discard the supernatant, add serum-containing sheep umbilical cord mesenchymal stem cell culture medium to the cell pellet, pipette evenly, and inoculate in a culture plate for cultivation;

[0031] (3) When the confluence of the cells cultured in step ...

Embodiment 2

[0035] In this embodiment, a Hu sheep semen cryopreservation solution contains the following components: Tris 35g / L, citric acid 17.5g / L, fructose 6g / L, sorbitol 20g / L, glycerin 5.5v / v%, BSA 4g / L, mesenchymal stem cell freeze-dried powder 0.5g / L, penicillin 1 million IU / L, streptomycin 1g / L.

[0036] The preparation method of the Hu sheep mesenchymal stem cell freeze-dried powder is the same as that in Example 1.

Embodiment 3

[0038] In this embodiment, a Hu sheep semen cryopreservation solution contains the following components: Tris 33g / L, citric acid 15.5g / L, fructose 9g / L, sorbitol 15g / L, glycerin 10v / v%, BSA 2.5 g / L, mesenchymal stem cell freeze-dried powder 0.4g / L, penicillin 1 million IU / L, streptomycin 1g / L.

[0039] The preparation method of the Hu sheep mesenchymal stem cell freeze-dried powder is the same as that in Example 1.

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PUM

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Abstract

The invention provides a freezing medium for semen of Hu sheep. The Hu sheep semen cryopreservation solution comprises the following components: 25-40 g / L of Tris, 15-18 g / L of citric acid, 5-15 g / L of fructose, 5-20 g / L of sorbitol, 3-10 v / v% of glycerol, 2-7 g / L of BSA (Bovine Serum Albumin) and 0.2-0.8 g / L of Hu sheep mesenchymal stem cell freeze-dried powder. The Hu sheep semen cryopreservation liquid can effectively reduce damage and'aging 'phenomena of Hu sheep semen in the cryopreservation process, and can well maintain sperm activity and acrosome integrity. When the cryopreservation liquid and the cryopreservation method are used for cryopreservation of the Hu sheep semen at the ultralow temperature, the unfrozen semen still has good insemination ability, the fetal effect is stable, the requirement for artificial insemination of the Hu sheep frozen semen can be met, and the cryopreservation liquid and the cryopreservation method have wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of Hu sheep breeding, and in particular relates to a Hu sheep semen freezing solution containing exosomes and a freezing method. Background technique [0002] Hu sheep is one of the important livestock in the Taihu Plain, and it is a first-class protected local livestock breed in my country. In animal husbandry classification, it is a rough-wool sheep that is used for both lamb skin and meat. Hu sheep have been bred and raised in the Taihu Plain for more than 800 years. Due to the natural conditions of Taiping Lake and the influence of human selection, a unique and rare breed was gradually bred. The production area is in the Taihu Lake Basin between Zhejiang and Jiangsu, so it is called "Hu sheep". [0003] In order to improve the reproductive efficiency of Hu sheep and make full use of the semen of good breeding males, artificial insemination technology has been applied to the breeding of Hu sheep in actu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/0775
CPCA01N1/0226A01N1/0221A01N1/021C12N5/0668C12N2509/10C12N2509/00
Inventor 王争光曾建露姚婷韩强张云超王邵雨陈曦
Owner ZHEJIANG UNIV
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