Semen diluting liquid capable of improving low-temperature/cryopreservation quality of semen and application of semen diluting liquid

A cryopreservation solution and cryopreservation technology, applied in the field of semen diluent, can solve the problems of decreased effective sperm count, low conception rate, and decreased sperm motility, and achieve the goals of improving the low-temperature preservation activity of semen, easy operation, and improved preservation quality Effect

Inactive Publication Date: 2017-10-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ultra-low temperature cryopreservation is usually achieved by storing semen in 0.5mL or 0.25mL plastic thin tubes. After thawing, the straight forward movement rate of sperm is about 45%. Using frozen semen for artificial insemination, equine conception The rate is significantly lower than that of fresh semen and liquid-preserved semen, because the vitality of the frozen and thawed semen is reduced, and the number of effective sperm that can maintain the fertilization ability is significantly reduced

Method used

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  • Semen diluting liquid capable of improving low-temperature/cryopreservation quality of semen and application of semen diluting liquid
  • Semen diluting liquid capable of improving low-temperature/cryopreservation quality of semen and application of semen diluting liquid
  • Semen diluting liquid capable of improving low-temperature/cryopreservation quality of semen and application of semen diluting liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Semen Diluent (for cryopreservation)

[0025] This example is used to illustrate the preparation of semen diluent for cryopreservation of semen.

[0026] 1. Ingredients: glucose, lactose, raffinose, sodium citrate, potassium citrate, HEPES reagent, BAPTA-AM, the above drugs are of analytical grade, distilled water, skimmed milk, penicillin, streptomycin.

[0027] 2. Preparation method: prepare 200mL of semen diluent as an example, first weigh 5.0g of glucose, 0.3g of lactose, 0.3g of raffinose, 0.06g of sodium citrate, 0.082g of potassium citrate, and 0.952g of HEPES reagent, and weigh them Put it into an Erlenmeyer flask, add 100mL of distilled water to mix, shake the Erlenmeyer flask until the solid is dissolved; pour the skimmed milk into another clean Erlenmeyer flask; put it into an autoclave for sterilization at 115°C for 20 minutes. After cooling at room temperature, mix the two, add 200,000 IU of penicillin and streptomycin and BAPTA-AM at a final con...

Embodiment 2

[0028] Example 2 Semen Diluent (for cryopreservation)

[0029] This example is used to illustrate the preparation of semen dilutions for semen cryopreservation.

[0030] 1. Ingredients: glucose, lactose, raffinose, sodium citrate, potassium citrate, HEPES reagent, BAPTA-AM, the above drugs are of analytical grade, distilled water, skim milk, penicillin, streptomycin, sterilized glycerin, egg yolk .

[0031] 2. Preparation method: Taking the preparation of 200mL as an example, first weigh 4.5g of glucose, 0.27g of lactose, 0.27g of raffinose, 0.054g of sodium citrate, 0.0738g of potassium citrate, and 0.8568g of HEPES reagent, and put them into Add 90mL of distilled water to the Erlenmeyer flask, shake the flask until the solid is dissolved; pour 90mL of skim milk into another clean Erlenmeyer flask; at the same time put it into an autoclave for sterilization, and sterilize at 115°C for 20min. After disinfection, take it out, and mix the two after cooling at room temperature,...

Embodiment 3

[0032] Example 3 Method for improving the quality of semen cryopreservation

[0033] 1. Manually collect horse semen, the semen collection device is CAU-H1 type semen collection device. Before sperm collection, add an appropriate amount (1000-1500ml) of warm water (42-45°C) into the false vagina. After the stallion's penis becomes erect, wash it with club soda, then rinse with water and dry. Apply lubricant evenly in the disposable lining (1 / 3 depth) before semen collection, and adjust the pressure of the sperm collection cylinder according to the length of the stallion's penis. After the semen collection device is ready, the semen collection officer stands on the right rear side of the horse, the trainer leads the stallion to climb over the false horse, and guides the penis into the semen collection tube. After the stallion ejaculates, insert the semen collection device The end of the collection cup is tilted downwards, and at the same time, the air release valve is opened ...

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Abstract

The invention relates to the field of farming, and concretely discloses a semen diluting liquid capable of improving low-temperature / cryopreservation quality of semen and application of the semen diluting liquid. The semen diluting liquid contains BAPTA-AM with a concentration of 40-60 [m]M, preferably 50 [m]M. The semen diluting liquid provided by the invention is capable of substantially improving the low-temperature preservation activity of semen and prolonging the in-vitro preservation time of semen. Meanwhile, the activity of semen subjected freezing and unfreezing is improved by 10%-20% compared with the activity of a control group. The application provided by the invention is simple to operate, the application cost is low, the effect is substantial, and no harmful influence is generated on animals. The semen diluting liquid can be widely applied to equid semen preservation and relevant embryonic implantation industry and embryonic engineering, and has a wide market prospect.

Description

technical field [0001] The invention relates to the field of animal husbandry, in particular to a semen diluent for improving the quality of semen cryopreservation / freezing and an application thereof. Background technique [0002] Semen cryopreservation technology can make full use of the genetic resources of fine breeding stock and rapidly expand the proportion of fine breeds in the herd. At the same time, this technology is also an effective auxiliary means for the treatment of infertility in mammals. Semen diluent is a protective agent for sperm, which has an important impact on the effect of semen preservation. The types of semen diluents commonly used in mammals include citric acid-sugar-yolk diluents, sucrose diluents with antioxidants, and milk diluents. Regardless of the type of semen diluents used, they should generally consist of osmotic pressure-maintaining substances, bacteriostatic substances, cryoprotectants, or cryoprotectants. Among them, the antifreeze pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0221A01N1/0226
Inventor 曾申明吴帅帅
Owner CHINA AGRI UNIV
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