Method for in vitro capacitation of rabbit sperms and nutrient solution formula

A culture medium and capacitation technology, applied in the field of biology, can solve the problems of reducing the effect of sperm energy and affecting the effect of in vitro fertilization, etc.

Inactive Publication Date: 2014-09-24
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the process of sperm capacitation in vitro, sperm will produce a variety of harmful substances, such as active oxygen and oxygen free radicals produced by ce...

Method used

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  • Method for in vitro capacitation of rabbit sperms and nutrient solution formula
  • Method for in vitro capacitation of rabbit sperms and nutrient solution formula
  • Method for in vitro capacitation of rabbit sperms and nutrient solution formula

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Experimental program
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Embodiment Construction

[0025] Technical thinking of the present invention is:

[0026] In order to avoid semen as the research object, by adding different concentrations of EGCG to the sperm BO capacitation solution, the effect of EGCG on sperm capacitation was studied in order to improve the effect of sperm capacitation in vitro, aiming to establish an efficient in vitro capacitation culture system for rabbits. Improve the corresponding embryo production efficiency.

[0027] (Note: BO fluid is a routinely used capacitation fluid).

[0028] The present invention will be specifically described below.

[0029] 1. Test animals

[0030] Healthy adult New Zealand male rabbits were selected as experimental rabbits.

[0031] 2. Experimental grouping

[0032] BO capacitation solution: NaCl 6.5453mg / mL, KCl 0.2997mg / mL, NaH 2 PO 4 ·H 2 O 0.1145 mg / mL, MgCl 2 ·6H 2 O 0.1057mg / mL, CaCl 2 2H 2 O0.330mg / mL, glucose 2.518mg / mL, 0.2% phenol red 4%, 100IU / mL penicillin, 100IU / mL streptomycin, NaHCO 3 3....

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Abstract

The invention relates to a method for in vitro capacitation of rabbit sperms and a nutrient solution formula. In the invention, 10 mum mol/L of EGCG is added in a BO capacitation liquid for sperm in vitro capacitation treatment, and 10 mum mol/L of EGCG is added in the BO capacitation liquid, the formula of the a BO capacitation liquid comprises the following components by weight: 6.5453mg/mL of NaCl, 0.2997mg/mL of KCL, 0.1145mg/mL of NaH2PO4.H2O, 0.1057mg/mL of MgC12.6H2O, 0.330mg/mL of CaCl2.2H2O, 2.518mg/mL of glucose, 4% of 0.2% phenol red, 100IU/mL of penicillin, 100IU/mL of streptomycin, 3.1080mg/mL of NaHCO3, 0.1380mg/mL of sodium pyruvate, 20mum g/mL of heparin and 20mg/mL of BSA. The method for in vitro capacitation of rabbit sperms overcomes the defect of mass oxyradical accumulation generated by sperm metabolism due to large sperm density during a current sperm in vitro capacitation process. The method uses adult New Zealand white male rabbit, different concentration EGCG (0mumM, 10mumM, 15mumM, 20mumM and 25mumM) can be added in the BO capacitation liquid, influence of the BO capacitation liquid to the rabbit sperm capacitation effect is researched so that optimum EGCG addition concentration is obtained, and thereby a rabbit in vitro capacitation culture system with high efficiency is established.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method for in vitro capacitation of rabbit sperm and a formula of culture solution. Background technique [0002] In vitro fertilization technology is an important part of in vitro embryo production. Through in vitro fertilization, a large number of high-quality and cheap embryos can be produced to meet the needs of embryo commercialization. Sperm capacitation in vitro is an essential part of in vitro fertilization technology. During sperm capacitation, a series of physiological and biochemical reactions occur inside the sperm, including the removal of seminal plasma proteins on the sperm surface and the movement of ions between the inside and outside of the cell membrane. (Ca 2+ , Mg 2+ , Cl - and HCO 3 - K + 、Na + efflux), opening of sperm capacitation regulatory system, hyperpolarization of sperm membrane potential, etc. The molecular mechanisms of sperm capacitation mainly in...

Claims

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Application Information

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IPC IPC(8): C12N5/076
Inventor 李拥军张光景张佳慧杨波蔡缪荧
Owner YANGZHOU UNIV
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