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Extremely-low sperm freezing protective agent and application thereof

A cryoprotectant and cryopreservation technology, which is applied in the application, preservation of human or animal body, animal husbandry, etc., can solve the problems of sperm damage, osmotic pressure local reactive oxygen species damage sperm structure and function, sperm apoptosis, etc. Achieving the effect of high freezing recovery rate

Active Publication Date: 2015-02-25
HAOHONG BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the small number and poor quality of rare sperm from various types clinically, especially the sperm of testicular tissue obtained through microsurgery, the movement of this part of sperm is very weak when observed under a microscope, and the sperm cell membrane They are not yet fully mature, and the stability of sperm nuclear DNA is also poor. If this part of sperm is frozen according to the traditional cryoprotectant method, factors such as temperature drop and intracellular ice crystal formation during the freezing process will cause a certain degree of sperm damage, such as The mechanical damage of cell structure, sharp change of osmotic pressure, and rapid increase of local active oxygen content caused by ice crystal formation can damage the structure and function of sperm plasma membrane, flagella, mitochondria, acrosome, nucleus and other organelles, affecting the survival rate of sperm , vitality, shape, fertilization ability and the integrity of the genetic material carried, and even induce sperm apoptosis

Method used

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  • Extremely-low sperm freezing protective agent and application thereof
  • Extremely-low sperm freezing protective agent and application thereof
  • Extremely-low sperm freezing protective agent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Cryoprotectant for rare sperm of the present invention (1)

[0029] Sodium chloride 5.5mmol / L, potassium chloride 4.5mmol / L, magnesium sulfate 0.2mmol / L, potassium dihydrogen phosphate 0.37mmol / L, calcium chloride 2.0mmol / L, fructose 3.0mmol / L, trehalose 3.0mmol / L, sodium citrate 1.37mmol / L, glycine 1mmol / L, glutamine 1mmol / L, pentoxifylline 2.0 mmol / L, vitamin E 3.0ml / L, HAS 2.0g / L, glycerin 140ml / L, Gentamicin sulfate 1.0g / L, the balance is ultrapure water.

[0030] Specific configuration process: Take a sterilized clean glass container and fill it with 500ml ultrapure water. Then use an analytical balance to accurately weigh each component successively, add sodium chloride, potassium chloride, magnesium sulfate, potassium dihydrogen phosphate, calcium chloride, fructose, trehalose, sodium citrate, glutamine, pentoxifylline , glycine, vitamin E, HAS, glycerin, gentamicin sulfate, fully stir and mix with a magnetic stirrer, and add ultrapure water to 1000...

Embodiment 2

[0031] Example 2 Cryoprotectant for rare sperm of the present invention (2)

[0032] Sodium chloride 5.8mmol / L, potassium chloride 4.2mmol / L, magnesium sulfate 0.22mmol / L, potassium dihydrogen phosphate 0.35mmol / L, calcium chloride 2.1mmol / L, fructose 2.8mmol / L, trehalose 3.2mmol / L, sodium citrate 1.35mmol / L, glycine 1.04mmol / L, glutamine 0.95mmol / L, pentoxifylline 2.1 mmol / L, vitamin E 2.8ml / L, HAS 2.2g / L, glycerol 130ml / L, gentamicin sulfate 1.1g / L, surplus is ultrapure water.

Embodiment 3

[0033] Example 3 Cryoprotectant for rare sperm of the present invention (3)

[0034]Sodium chloride 5.2mmol / L, potassium chloride 4.6mmol / L, magnesium sulfate 0.18mmol / L, potassium dihydrogen phosphate 0.40mmol / L, calcium chloride 1.8mmol / L, fructose 3.2mmol / L, trehalose 2.8mmol / L, Sodium Citrate 1.40mmol / L, Glycine 0.92mmol / L, Glutamine 1.05mmol / L, Pentoxifylline 1.8mmol / L, Vitamin E 3.2ml / L, HAS 1.8g / L, Glycerin 150ml / L L, gentamicin sulfate 0.95g / L, and the balance is ultrapure water.

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Abstract

The invention relates to an extremely-low sperm freezing protective agent and application thereof. The protective agent is prepared from the following ingredients: sodium chloride, potassium chloride, magnesium sulfate, monopotassium phosphate, calcium chloride, fructose, trehalose, sodium citrate, glycin, glutamine, pentoxifylline, vitamin E, HAS (human serum albumin), glycerin and gentamicin sulphate. The extremely-low sperm freezing protective agent can protect less sperms with low activity and without forward activity ability in seminal fluid, can remarkably improve the activity ability of the sperms which swing weakly and cannot move forwards in seminal fluid after freezing and thawing, has no obvious influence on the physiological function of the sperms, has high freezing anabiosis rate, can be used for cryopreservation of less and weak sperms, testicle sperms and epididymis sperms, and contributes to the successful development of assisted reproductive technology.

Description

technical field [0001] The invention relates to the technical field of sperm preservation, in particular to a rare sperm cryoprotectant and its application. Background technique [0002] With the advancement of modern civilization and the development of social industrialization, environmental pollution has become more and more serious, and semen has increased abnormally, causing the incidence of infertility to reach 10%, becoming a common disease. In 1992, the emergence of intracytoplasmic sperm injection (ICSI) technology brought good news to infertile patients. Patients with severe male infertility, such as severe oligozoospermia or asthenozoospermia, that is, sperm density < (1-5) × 10 6 / mL (1 to 5 strips per high power field (HPF)), patients with obstructive azoospermia and non-obstructive azoospermia can obtain sperm from the testis or epididymis through surgery, although the number of sperm obtained is small, However, they can all have offspring through ICSI tech...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 刘锋高一丰盖安生
Owner HAOHONG BIOTECH SHANGHAI
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