A method for capacitation of yak sperm in vitro

A technology of sperm capacitation and sperm, which is applied in the field of yak breeding, can solve the problems of unstable results, poor repeatability, unstable cleavage rate of fertilized eggs and in vitro development rate of blastocysts, etc., and achieves convenient operation, easy production and use, and measurement The results are accurate

Inactive Publication Date: 2016-05-11
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] With the development of modern biotechnology, embryo biotechnology is gradually applied to yak scientific research, but due to poor repeatability and unstable results, yak embryo biotechnology has not been effectively carried out in yak production
Especially in yak in vitro fertilization technology, the common problem is that the cleavage rate of fertilized eggs and the in vitro development rate of blastocysts are unstable, and the overall success rate of in vitro fertilization is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] One vial of frozen yak semen was thawed for in vitro capacitation.

[0020] (1) Take out a thin tube of yak frozen semen from the liquid nitrogen tank, stay in the air for 6 seconds, then put it into a constant temperature water bath at 37°C to 38°C, and take it out when 1 / 2 of it melts.

[0021] (2) Use sterilized toilet paper to wipe off the water outside the thin tube, then quickly wipe it once with an alcohol cotton ball, and place it in an ultra-clean workbench to dissolve it completely.

[0022] (3) Sperm capacitation was treated with the improved TALP liquid flotation method; the steps of the flotation method were: add 1 mL of sperm capacitation solution to the dissolved semen, mix well, and divide into 4 small plastic centrifuge tubes, 38.5 ℃ CO 2 Float in the incubator for 1 hour, then carefully collect the supernatant from the 4 centrifuge tubes into a 10mL centrifuge tube, about 0.6-0.8mL, centrifuge and wash twice at 700g, 10min each time; / mL heparinized s...

Embodiment 2

[0026] Two vials of yak frozen semen were thawed for in vitro capacitation.

[0027] (1) Take out two thin tubes of yak frozen semen from the liquid nitrogen tank, stay in the air for 6 seconds, then put them into a constant temperature water bath at 37°C to 38°C, and take them out when 1 / 2 of them melt.

[0028] (2) Use sterilized toilet paper to wipe off the water outside the thin tube, then quickly wipe it once with an alcohol cotton ball, and place it in an ultra-clean workbench to dissolve it completely.

[0029] (3) Sperm capacitation was treated with the improved TALP liquid flotation method; the steps of the flotation method were: add 1.5mL sperm capacitation liquid to the dissolved semen, mix well and divide into 6 small plastic centrifuge tubes at 38.5°C CO 2 Float in the incubator for 1 hour, then carefully collect the supernatant from 6 centrifuge tubes into a 15mL centrifuge tube, about 0.8-1.2mL, centrifuge and wash twice at 800g, 8min each time; / mL heparinize...

Embodiment 3

[0033] 200 yak oocytes were fertilized in vitro with frozen semen for external capacitation.

[0034] (1) Take out two thin tubes of yak frozen semen from the liquid nitrogen tank, stay in the air for 6 seconds, then put them into a constant temperature water bath at 37°C to 38°C, and take them out when 1 / 2 of them melt.

[0035] (2) Use sterilized toilet paper to wipe off the water outside the thin tube, then quickly wipe it once with an alcohol cotton ball, and place it in an ultra-clean workbench to dissolve it completely.

[0036] (3) Sperm capacitation was treated with the improved TALP liquid flotation method; the steps of the flotation method were: add 1.5mL sperm capacitation liquid to the dissolved semen, mix well and divide into 6 small plastic centrifuge tubes at 38.5°C CO 2 Float in the incubator for 1 hour, then carefully collect the supernatant from 6 centrifuge tubes into a 15mL centrifuge tube, about 0.8-1.2mL, centrifuge and wash twice at 700-800g, 8-10min / ti...

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Abstract

The invention discloses a method for in vitro capacitation of yak sperms. The method mainly comprises a method for unfreezing yak frozen semen, sperm in vitro capacitation liquid, a method for capacitation, sperm in vitro fertilization liquid and a method for evaluating the capacitation effect. By adopting the method, a specific condition for in vitro capacitation of the yak sperms is provided to ensure the quality and efficiency of the in vitro capacitation of the yak sperms. The method is comprehensive in technology and method, convenient to operate and accurate in determination result, can be completely used for in vitro capacitation of the yak sperms or in vitro production of embryos, and can effectively improve the breeding efficiency of yaks.

Description

technical field [0001] The invention belongs to the field of yak reproduction, and relates to yak embryo biotechnology, in particular to a method for capacitation of yak sperm in vitro. Background technique [0002] Yak is a unique cattle species distributed in the Qinghai-Tibet Plateau and adjacent areas. The reproduction of yak has obvious seasonality, and the estrus of yak has the general characteristics of ordinary cattle, but it is not as obvious as that of ordinary cattle. However, due to the special natural ecological environment conditions, the yak has the ecological and physiological characteristics of late maturity and low fecundity. [0003] With the development of modern biotechnology, embryo biotechnology has been gradually applied to yak scientific research, but due to poor repeatability and unstable results, yak embryo biotechnology has not been effectively carried out in yak production. Especially in yak in vitro fertilization technology, the ubiquitous pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/076C12Q1/02
Inventor 郭宪丁学智阎萍梁春年王宏博
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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