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Sperm freezing medium as well as preparation method and application thereof

A technology of cryopreservation and preservation method, applied in the application, preservation of human or animal body, animal husbandry, etc., can solve the problem of no oxidative stress, achieve prevention of oxidative stress, high quality, and eliminate potential allergic reactions Effect

Active Publication Date: 2019-02-15
太东(镇江)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

while current products offer little to no measure against oxidative stress

Method used

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  • Sperm freezing medium as well as preparation method and application thereof
  • Sperm freezing medium as well as preparation method and application thereof
  • Sperm freezing medium as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Preparation of cryopreservation solution (take 10L as an example)

[0039] A. Prepare according to the components in Table 1, prepare the solid first and then the solution, set the volume to 10L and filter with 0.22μm.

[0040] Table 1 Components

[0041]

[0042]

[0043] a. Accurately weigh the solid components of potassium chloride, magnesium sulfate, calcium chloride, sodium bicarbonate, dipotassium hydrogen phosphate, sodium pyruvate, calcium lactate, glucose, sucrose and HEPES, and dissolve them with ultrapure water for injection.

[0044] b. Filter the above liquid through a 0.22 μm filter membrane.

[0045] c. Add glutamine, glycine, gentamicin, glycerin, vitamin E, vitamin C and vitamin B12 in the components, measure the osmotic pressure and pH value of the solution, at this time, the osmotic pressure of the solution is 580-600mOsm / kg , pH value is 7.0-7.4.

[0046] d. Add recombinant human serum albumin so that the final albumin concentration is 5...

experiment example 1

[0087] Adopt embodiment 1 and the cryopreservation liquid that comparative example 1-3 makes to carry out cryopreservation test, specifically as follows:

[0088] a. Add sperm liquid (about 0.5ml) to 1ml freezing solution and let it stand at room temperature for 5 minutes;

[0089] b. Place the cryovial at 4°C for 5 minutes;

[0090] c. Place the frozen tube at -20°C for 5 minutes;

[0091] d. Place the cryopreservation tube at -80°C for 5 minutes;

[0092] e. Cryopreservation tubes are stored in liquid nitrogen.

[0093] After storage in liquid nitrogen for a period of time, resuscitate according to the following steps:

[0094] After the cryopreservation tube was taken out from the liquid nitrogen, it was shaken and rewarmed at 37°C for 2 minutes, and then washed with washing solution.

[0095] After thawing, the proportion of forward motile sperm was counted again.

[0096] In this experiment example, the selected sperm density is 40-80×10 6 pcs / ml, the activity rate ...

experiment example 2

[0103] The quality of sperm before cryopreservation and after recovery from cryopreservation for 1 h was tested, and the normal rate of sperm morphology was tested. The results are shown in Table 3.

[0104] Table 3 Sperm quality

[0105]

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PUM

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Abstract

The invention relates to the field of sperm freezing, and particularly relates to a sperm freezing medium as well as a preparation method and application thereof. The sperm freezing medium comprises the following components: potassium chloride, magnesium sulfate, calcium chloride, sodium bicarbonate, dipotassium phosphate, sodium pyruvate, calcium lactate, glucose, saccharose, glutamine, glycine,HEPES, gentamicin, glycerinum, human serum albumin, vitamin E, vitamin C and vitamin B12. According to the sperm freezing medium, all components are matched with each other to comprehensively maintainthe osmotic pressure, the straining capacity of sperm to resist the outside environment change; the freezing medium is used for storing sperm in low temperature; the sperm survive rate and the activity can be maintained for a long term.

Description

technical field [0001] The invention relates to the field of sperm cryopreservation, in particular to a sperm cryopreservation solution and its preparation method and application. Background technique [0002] Human sperm cryopreservation is very important for human reproductive health. Those who are unwilling to bear children in the short term can freeze viable sperm when they are young, and perform artificial insemination after thawing when needed. Or patients who are going to receive radiotherapy or chemotherapy, or are about to enter work that is harmful to reproductive health, can freeze their sperm so that they can obtain healthy offspring in the future. [0003] The process of sperm cryopreservation is mainly to gradually cool down the temperature so that the sperm are finally frozen in liquid nitrogen. The ice crystals generated during the cryopreservation process will greatly damage the sperm cells. The function of the sperm freezing solution is to protect the spe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 李兵
Owner 太东(镇江)生物科技有限公司
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