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A kind of bovine sperm capacitation liquid and sperm capacitation method in vitro

A technology of sperm capacitation and capacitation, which is applied in the field of livestock breeding, can solve problems such as easy resistance to antibiotics, and achieve the effects of avoiding drug resistance, improving production quality and efficiency, and simple operation

Inactive Publication Date: 2019-09-03
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two capacitation solutions contain antibiotics penicillin and streptomycin, and antibiotics are prone to resistance problems, and the side effects caused by them cannot be ignored.

Method used

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  • A kind of bovine sperm capacitation liquid and sperm capacitation method in vitro
  • A kind of bovine sperm capacitation liquid and sperm capacitation method in vitro
  • A kind of bovine sperm capacitation liquid and sperm capacitation method in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Bovine sperm capacitation fluid, consisting of the following components: NaCl 6.7500mg / mL, KCl 0.3000mg / mL, NaHCO 3 2.5000mg / mL, NaH 2 PO 4 ·2H 2 O 0.0620mg / mL, CaCl 2 ·2H 2 O 0.2960 mg / mL, MgCl 2 ·6H 2 O0.1000mg / mL, sodium pyruvate 0.1120mg / mL, EDTA-Na 0.0186mg / mL, phenol red 0.0100mg / mL, BSA 6.0000mg / mL, glucose 1.0000mg / mL, caffeine 0.4800mg / mL, heparin 0.0500 mg / mL, nisin 0.025mg / mL, and the balance is sub-boiling water for solvent, pH 7.5.

[0023] The method for in vitro capacitation of bovine sperm includes the following steps:

[0024] 1) Take out the frozen Holstein cattle essence from the liquid nitrogen tank, and quickly put it into a 38°C water bath to thaw it for 15s;

[0025] 2) Slowly add the thawed semen to the preheated balance (at 38°C, 5% CO 2 , saturated humidity CO 2 After preheating and equilibrating for 1 h) in the incubator, the bottom of the capacitative solution test tube was inclined at an angle of 45°, and placed in 38°C, 5% CO 2 ,...

Embodiment 2

[0027] Bovine sperm capacitation fluid, consisting of the following components: NaCl 6.7500mg / mL, KCl 0.3000mg / mL, NaHCO 3 2.5000mg / mL, NaH 2 PO 4 ·2H 2 O 0.0620mg / mL, CaCl 2 ·2H 2 O 0.2960 mg / mL, MgCl 2 ·6H 2 O0.1000mg / mL, sodium pyruvate 0.1120mg / mL, EDTA-Na 0.0186mg / mL, phenol red 0.0100mg / mL, BSA 6.0000mg / mL, glucose 1.0000mg / mL, caffeine 0.4800mg / mL, heparin 0.0500 mg / mL, nisin 0.0375mg / mL, and the balance is sub-boiling water, pH 7.6.

[0028] The method for in vitro capacitation of bovine sperm includes the following steps:

[0029]1) Take out the frozen Holstein beef semen from the liquid nitrogen tank, and quickly put it into a 38°C water bath to thaw for 10 seconds;

[0030] 2) Slowly add the thawed semen to the preheated equilibrium (at 39°C, 5% CO 2 , saturated humidity CO 2 The bottom of the capacitation solution test tube after preheating and equilibrating in the incubator for 1.5h) is tilted at an angle of 45°, and placed in 38°C, 5% CO 2 , saturated ...

Embodiment 3

[0032] Bovine sperm capacitation solution, consisting of the following components: NaCl 6.7500mg / mL, KCl 0.3000mg / mL, NaHCO 3 2.5000mg / mL, NaH 2 PO 4 2H 2 O 0.0620mg / mL, CaCl 2 2H 2 O 0.2960 mg / mL, MgCl 2 ·6H 2 O0.1000mg / mL, sodium pyruvate 0.1120mg / mL, EDTA-Na 0.0186mg / mL, phenol red 0.0100mg / mL, BSA6.0000mg / mL, glucose 1.0000mg / mL, caffeine 0.4800mg / mL, heparin 0.0500 mg / mL, nisin 0.05mg / mL, the balance is sub-boiling water, pH7.8.

[0033] A bovine sperm capacitation method in vitro, comprising the following steps:

[0034] 1) Take out the frozen Holstein beef semen from the liquid nitrogen tank, and quickly put it into a 40°C water bath to thaw for 10 seconds;

[0035] 2) Slowly add the thawed semen to the preheated equilibrium (at 38.5°C, 5% CO 2 , saturated humidity CO 2 The bottom of the capacitation solution test tube after preheating and equilibrating in the incubator for 2h) is tilted at an angle of 45°, and placed in 38.5°C, 5% CO 2 , saturated humidity C...

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Abstract

Belonging to the technical field of livestock breeding, the invention discloses a bovine sperm capacitation solution and an in vitro sperm capacitation method. The bovine sperm capacitation solution mainly consists of the following components: 6.7500mg / mL NaCl, 0.3000mg / mL KCl, 2.5000mg / mL NaHCO3, 0.0620mg / mL NaH2PO4.2H2O, 0.2960mg / mL CaCl2.2H2O, 0.1000mg / mL MgCl2.6H2O, 0.1120mg / mL sodium pyruvate, 0.0186mg / mL EDTA-Na, 0.0100mg / mL phenol red, 6.0000mg / mL BSA, 1.0000mg / mL glucose, 0.4800mg / mL caffeine, 0.0500mg / mL heparin, and 0.025-0.05mg / mL Nisin. The capacitation solution provided by the invention can effectively ensure the sperm motility, sperm plasma membrane integrity rate, acrosomal integrity rate and mitochondrial activity, and maintains the in vitro sperm capacitation effect.

Description

technical field [0001] The invention relates to a bovine sperm capacitation liquid and a sperm capacitation method in vitro, which belongs to the technical field of livestock reproduction. Background technique [0002] With the development of artificial insemination and in vitro fertilization technology, the fertilization ability of bulls has become an important factor affecting the reproductive efficiency of cattle (Hoflack et al., 2006). While semen quality analysis is a common procedural tool for assessing bull fertility potential (Rijsselaere et al., 2002), bovine semen quality analysis is particularly important (Hoflack et al., 2007). At the same time, the quality of semen is also closely related to the conception rate of cows, which is a key factor to improve the reproductive efficiency of cattle. There are many factors that affect bull semen quality, such as age, nutrition, season and capacitation effect. The effective premise of ensuring sperm capacitation is addin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/076
Inventor 李晓霞曹平华文凤云许常龙孙俊丽禹学礼栗颖华付诗卉韩文霞武花
Owner HENAN UNIV OF SCI & TECH
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