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In Vitro Maturation Culture Medium of Small Follicle Oocytes and Its Application

A technology for in vitro maturation and culture of oocytes, applied in the field of in vitro maturation culture medium, can solve the problems of not improving the developmental ability of oocytes, and achieve the effects of improving in vitro developmental ability, less toxic effect, and promoting synchronous cytoplasmic maturation

Active Publication Date: 2021-05-14
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in simulating the in vivo environment, a two-stage maturation method of oocytes in vitro was developed, and the research results showed that meiosis inhibitors did not improve the ability of oocytes to develop in vitro, and even had adverse effects

Method used

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  • In Vitro Maturation Culture Medium of Small Follicle Oocytes and Its Application
  • In Vitro Maturation Culture Medium of Small Follicle Oocytes and Its Application
  • In Vitro Maturation Culture Medium of Small Follicle Oocytes and Its Application

Examples

Experimental program
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Effect test

Embodiment 1

[0038] The in vitro maturation medium of embodiment 1 small follicle oocyte

[0039] 1. Raw materials:

[0040] TCM-199, BSA, FSH, E2, BNP and / or CNP, cysteine ​​(Cys), penicillin, streptomycin.

[0041]2. Preparation method:

[0042] Dissolve BSA, FSH, E2, BNP / CNP, cysteine, penicillin, and streptomycin in TCM-199 to prepare 4mg / mL BSA, 0.05IU / mL FSH, 100nM E2, 100ng / mL BNP, 0.1mg / mL cysteine ​​(Cys), 100IU / mL penicillin, 100IU / mL streptomycin TCM-199 culture medium is the culture medium for in vitro maturation.

Embodiment 2

[0043] The in vitro maturation medium of embodiment 2 small follicle oocyte

[0044] The difference between this embodiment and Embodiment 1 is that: BNP is replaced by CNP.

Embodiment 3

[0045] The in vitro maturation medium of embodiment 3 small follicle oocyte

[0046] The difference between this example and Example 1 is that the concentration of E2 is 120 nM, and the concentration of BNP is 150 ng / mL.

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Abstract

The invention relates to cell culture, and specifically discloses an in vitro maturation culture medium of small follicle oocytes, which contains 100nM E2 and 100ng / mL CNP / BNP. The invention also provides the application of the in vitro maturation culture solution in promoting the in vitro maturation of small follicle oocytes or inhibiting the meiosis of small follicle oocytes and a culture method for promoting the in vitro maturation of oocytes, specifically: collecting Animal oocytes are classified according to the diameter of the follicle: the small follicle oocytes are placed in the aforementioned in vitro maturation medium for pre-mature culture for 20h, and then cultured in the maturation medium for 46±2h; the large follicle Oocytes were directly cultured in maturation medium for 46±2h. The invention adopts a two-stage in vitro maturation culture method, applies CNP / BNP and E2 pre-maturation to treat small follicle oocytes, promotes the synchronization of nuclear maturation and cytoplasmic maturation, improves in vitro development ability, and can become a new type of meiosis-inhibiting drug. Popularization and application in animal husbandry.

Description

technical field [0001] The present invention relates to cell culture, in particular to the in vitro maturation culture solution of small follicle oocytes. Background technique [0002] In livestock breeding, in vitro maturation of unstimulated ovary-derived oocytes is widely used to produce animal embryos. Therefore, in vitro oocyte maturation is an important platform technology for breeding, cloning and transgenic animal production. [0003] At present, the number of high-quality breeding males is small, the breeding speed is slow, and the production performance is backward, which are the key factors restricting the development of animal husbandry in my country. The cell number of in vitro fertilized embryos is significantly lower than that of in vivo embryos, which reflects that the in vitro culture system of oocytes is not ideal. The quality of in vitro fertilized embryos is an important factor affecting the pregnancy rate of embryo transfer and the survival rate of offs...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/32C12N2501/11C12N2501/30C12N2501/31C12N2501/392
Inventor 夏国良张延浩王超
Owner CHINA AGRI UNIV
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