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A method for delaying the aging of eggs cultured in vitro

An in vitro culture and egg technology, which is applied in the field of delaying the aging of in vitro cultured eggs, can solve the problems of the mechanism of improving egg aging without astaxanthin, so as to ensure the ability of in vitro development, delay or reverse the aging of eggs, and improve the increase of early apoptosis rate Effect

Active Publication Date: 2022-03-04
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no international research on the mechanism of astaxanthin improving egg aging

Method used

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  • A method for delaying the aging of eggs cultured in vitro
  • A method for delaying the aging of eggs cultured in vitro
  • A method for delaying the aging of eggs cultured in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Collection of mouse eggs

[0031] The ICR mice used in this experiment were purchased from the SPF-level experimental animal breeding room from Inner Mongolia University Experimental Animal Research Center. Each mouse was subcutaneously injected with 10 IU PMSG, 46-48 hours of intraperitoneal injection of 10 Iu HCG method, in HCG injection 14-16h After the cervical spine dislocation method, mice were separated from the back and cut the fallopian tube, and the falloplate was placed in the mouse operating liquid M2, and the eggs were removed from the eggs in the in vitro operating M2. The formula of the mouse body fluid is as shown in Table 1.

[0032] Table 1 mice outside octaves M2

[0033]

[0034]

Embodiment 2

[0035] Example 2 mouse pestic cells in vitro culture

[0036]The ovate-oocyte complex (COCS) of the removal MII egg (COCS) was placed in CZB culture solution (shown in Table 2) in vitro culture (6h, 12h, 18h, 24h, each microphone) 20-25 The egg is cultured in the condition of the pH of 7.2-7.4, the carbon dioxide concentration of 5%, the humidity of 100%, and the temperature of 37 ° C. After culturing different times in vitro, in vitro fertilization experiments.

[0037] Table 2 CZB culture solution (CHATOT-Ziomet-Bavister)

[0038]

Embodiment 3

[0039] Example 3 Mouse ovary cell extractive fertilization

[0040] Preparation of sperm: Biomei, selected sexual maturity (> 8 weeks), has proven to have fertilization capabilities through mating experiments, and the cervical spine is dead, cut the abdominal cavity, cut the epididymis tail, remove fat, use tweezers and needles , The needle punctures the epidermis epidermis and extruded the sperm of the aposenticone.

[0041] Sperm can be able to collect sperm groups immediately, placed in a balanced well-equilibrium, slightly swaying several times, making the sperm group dispersed, placed in pH 7.2-7.4, carbon dioxide concentration 5%, The humidity was 100%, and the temperature of 37 ° C was incubated for 1.5 h.

[0042] In vitro fertilization: Egg CoCs cultured in vitro (T6 + 20) in vitro (T6 + 20), add the amount of volume of the material to the sperm density in 1 × 10 6 Around, it was placed in a pH 7.2-7.4, the carbon dioxide concentration of 5%, the humidity was 100%, and th...

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Abstract

The invention provides a method for delaying the aging of eggs cultured in vitro, which belongs to the technical field of animal reproduction. The present invention finds that adding astaxanthin (Astaxanthin) during in vitro culture of eggs in culture solution can effectively improve the early embryonic development of aging eggs, increase the membrane potential of aging eggs, and reduce the apoptosis rate of aging eggs. The method of adding astaxanthin in the process of culturing eggs in vitro significantly slows down the aging speed of eggs cultured in vitro, improves the early embryonic development rate of aging eggs, and improves the development rate of aging eggs compared with conventional culture medium. 15%, able to provide high-quality eggs and early embryos for in vitro fertilization-embryo transfer (IVF-ET) technology.

Description

Technical field [0001] The present invention relates to a method of delaying in vitro culture of egg aging, belonging to the field of animal breeding technology. Background technique [0002] The reproductive ability of mammals is affected by a variety of factors, and the aging of the animal's own, aging of germ cells severely affects the reproductive efficiency of animals, and the aging of the egg has severely reduced the reproductive ability of female animals. The aging of the egg is related to many factors, age, oval cells, active oxygen increases, mitochondrial dysfunction, etc., can cause aging of eggs. Among them, oxidative stress results in an increase in active oxygen is a very important factor. When the egg is aging, the oxidation and antioxidant balance in the cells at this time tends to the oxidation state, thereby exciting the main components such as proteins and lipids. Similarly, due to various reasons to induce the oxidation stress, the age of cells will be acceler...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075C12N5/073
Inventor 梁成光温鑫韩哲程建响赵庭何东东杨旻昊
Owner INNER MONGOLIA UNIVERSITY
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