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Composition for promoting fertilization of frozen semen of mouse

A technology for freezing semen and composition, applied in application, microorganism, animal cells, etc., can solve the problems of poor recovery and fertilization efficiency of frozen-thawed sperm, failure to meet the requirements of seed preservation, etc.

Inactive Publication Date: 2020-05-08
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Sperm freezing technology is convenient, fast, and does not require special equipment. Its application prospects are quite broad. However, according to the results of relevant laboratories at home and abroad, the efficiency of sperm freezing and recovery in mice is closely related to the genetic background. The inbred mouse C57BL / 6J The recovery and fertilization efficiency of frozen-thawed sperm is the worst. Without the use of assisted fertilization technology, it is almost impossible to meet the conservation requirements

Method used

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  • Composition for promoting fertilization of frozen semen of mouse
  • Composition for promoting fertilization of frozen semen of mouse
  • Composition for promoting fertilization of frozen semen of mouse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1 A kind of sperm capacitation liquid

[0055] 101.6mM NaCl; 4.7mM KCl; 0.4mM KH 2 PO 4 ; 0.2mM MgSO 4 ·7H 2 O; 25mM NaHCO 3 ; 2mM CaCl 2 2H 2 O; 0.01mg / ml PVA (polyvinyl alcohol); 2.8mM glucose (Glucose); 0.3mM sodium pyruvate (Na pyruvate); 23.3mM 60% sodium lactate (60% Na Lactate); Sulfonic acid (Hypotaurine); 0.1mg / ml penicillin G (PenicillinG·K salt); 0.75mM MBCD (Methyl-β-cyclodextrin methyl-β-cyclodextrin), the pH value is 7.34±0.10.

Embodiment 2

[0056] Embodiment 2 A kind of sperm capacitation liquid

[0057] 120mM NaCl; 6.0mM KCl; 0.5mM KH 2 PO 4 ; 0.3mM MgSO 4 ·7H 2 O; 30mM NaHCO 3 ;5mM CaCl 2 2H 2 O; 0.05mg / ml PVA (polyvinyl alcohol); 3.5mM glucose (Glucose); 0.3mM sodium pyruvate (Na pyruvate); 25mM 60% sodium lactate (60% NaLactate); 0.2mg / ml hypotaurine (Hypotaurine); 0.2mg / ml penicillin G (PenicillinG · K salt); 0.75mM MBCD (Methyl-β-cyclodextrin), pH value is 7.34±0.10.

Embodiment 3

[0058] Embodiment 3 A kind of sperm capacitation liquid

[0059] 100mM NaCl; 4.0mM KCl; 0.2mM KH 2 PO4 ; 0.1mM MgSO 4 ·7H 2 O; 20mM NaHCO 3 ; 1mM CaCl 2 2H 2 O; 0.05mg / ml PVA (polyvinyl alcohol polyvinyl alcohol); 2mM glucose (Glucose); 01mM sodium pyruvate (Na pyruvate); 20mM 60% sodium lactate (60%Na Lactate); 0.1mg / ml hypotaurine ( Hypotaurine); 0.1mg / ml penicillin G (PenicillinG·K salt); 0.5mM MBCD (Methyl-β-cyclodextrin), pH value is 7.34±0.10.

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Abstract

The invention discloses a composition for promoting fertilization of frozen semen of a mouse. 0.01 mg / ml PVA and 0.75 mM methyl-beta-cyclic dextrin (MBCD) are added to an m HTF solution to serve as asperm capacitation solution; and 1.0 mM reduced glutathione is added to an m HTF solution to serve as a sperm in-vitro fertilization solution. The sperm capacitation solution used in the invention canremarkably protect the integrity of a plasma membrane and an acrosome, reduce the influence of ROS, and prominently improve survival rate and mobility of a sperm; and in combination with the in-vitrofertilization solution containing the reduced glutathione in the invention, an in-vitro fertilization success rate of a frozen-thawed sperm of a genetically modified mouse with an inbred strain C57BL / 6J as a background can be improved remarkably, and a two-cell developmental rate after fertilization is 60-65%.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a composition for promoting the fertilization of mouse frozen semen. Background technique [0002] Laboratory mice have been used as experimental animals since the 18th century, and are currently the most widely used, most clearly studied and most in-depth experimental animals. Mice have a short growth period, relatively easy to reproduce and maintain, and the correspondence rate between the mouse genome and the corresponding genes in the human genome is also above 98%. With the breakthroughs in the field of genetic modification technology in recent years, the development and application of gene editing technology, the number of disease model mice constructed by genetic modification is increasing at an alarming rate. Animal models of these diseases have become an important tool for researchers to study the mechanism of disease, prevention and treatment measures, as well as the ...

Claims

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Application Information

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IPC IPC(8): C12N5/076C12N15/877
CPCC12N5/061C12N15/8775C12N2500/50C12N2501/90C12N2501/999C12N2500/34C12N2500/12C12N2500/14C12N2500/16C12N2500/32
Inventor 高赛飞蔡卫斌李慧杨镇宇
Owner SUN YAT SEN UNIV
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