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Human-assisted reproduction method for non-human mammals

A non-human mammal, artificially assisted technology, applied in the field of artificial assisted reproduction of non-human mammals, can solve problems such as unreliability

Active Publication Date: 2022-01-18
吉林省奥金斯农牧科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Practice has proved that this indicator is not reliable

Method used

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  • Human-assisted reproduction method for non-human mammals
  • Human-assisted reproduction method for non-human mammals
  • Human-assisted reproduction method for non-human mammals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Retrieval and in vitro maturation of non-human mammalian oocytes.

[0041] (1) Obtaining non-human mammalian oocytes.

[0042] The ovaries of adult cattle were collected from the slaughterhouse, placed in 30°C normal saline and sent to the laboratory within 4 hours, washed three times in 37°C PBS solution, and then the follicles with a diameter of 2-8mm were extracted with a needle with a diameter of 0.7mm. Pick cumulus-nonhuman mammalian oocyte complexes (COCs) with uniform shape and compact structure under a stereoscope.

[0043] (2) Maturation of non-human mammalian oocytes.

Embodiment 2

[0045] Example 2: Detection and verification test of the quality of non-human mammalian oocytes after maturation.

[0046] 1. Determination of the membrane potential of non-human mammalian oocytes.

[0047] S1: Preparation of intracellular microelectrodes, select domestic glass microelectrode blanks (outer diameter 1.6 mm), use WLQ-2 automatic multi-tube microelectrode puller to draw microglass tubes with a tip diameter of ≤1 μm, and fill them with 1mol / L KCl solution, the electrode resistance is 10-20MΩ.

[0048] S2: Preparation of non-human mammalian oocyte fixation tube, first draw a thin glass tube with an outer diameter of 1.3mm and a length of 120mm into a tapered glass needle, then cut it off with a calciner and burn the section until it is blunt, A fixed straw with an inner diameter of 30-40 μm and an outer diameter of 100-200 μm is made.

[0049] S3: Determination of the membrane potential of non-human mammalian oocytes. On an inverted microscope (NIKON ET2000 type...

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Abstract

The invention provides a human-assisted reproduction method for non-human mammals. The method comprises the step of judging quality of in-vitro mature non-human mammal oocytes according to a non-human mammal oocyte membrane potential value X. Experiments show that non-human mammal oocytes with different membrane potentials have significant difference in subsequent development potential, so that the membrane potentials of mature non-human mammal oocytes have correlation with the maturation quality and subsequent development potentials of the non-human mammal oocytes. The membrane potentials of the mature non-human mammal oocytes can be used as an important judgment index for the maturation quality and subsequent development potentials of the non-human mammal oocytes. If a membrane potential value is negative and between-30 mV and-10 mV, bovine oocytes with the membrane potential value have the best quality and the highest development rate of subsequent fertilization development and nuclear transfer embryos.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an artificial assisted reproductive method for non-human mammals. Background technique [0002] The maturation of non-human mammalian oocytes cultured in vitro is a very important technical link in animal artificial assisted reproductive technology. With the research and application of embryo engineering technologies such as in vitro fertilization and nuclear transfer becoming more and more widely The quality of cell culture in vitro is one of the key prerequisites for determining the success or failure of embryonic bioengineering research and application. [0003] During zygote formation, both the sperm and the egg cell contribute genetic material. In addition, egg cells provide a large amount of essential nutrients for the development of early embryos. Therefore, as the basis for the development of fertilized eggs, the quality of non-human mammalian oocytes is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/873C12N5/073A01K67/027G01N15/10
CPCC12N15/873C12N5/0604A01K67/0273A01K67/027G01N15/1031A01K2227/101A01K2267/02G01N2015/1006
Inventor 丁方荣王海平王剑
Owner 吉林省奥金斯农牧科技发展有限公司