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Porcine oocyte in-vitro maturation culture solution and application thereof

A technology for in vitro maturation and culture of oocytes, applied in the field of porcine oocyte maturation culture medium in vitro, can solve problems such as infection, potential safety hazards, low concentration, etc., achieve high cleavage rate and blastocyst rate, stable properties, and Ingredient-specific effects

Active Publication Date: 2020-08-18
广东中芯种业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Secondly, porcine follicular fluid is also a potential source of disease infection, especially in the current situation of African swine fever, the use of follicular fluid from slaughterhouses has great safety risks
Even if African swine fever virus was not detected before use, it may be due to low concentration
Once the embryo is transferred to the recipient sow, the virus multiplies in the body, leading to infection of the recipient pig or surrounding pigs, and the loss will be huge

Method used

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  • Porcine oocyte in-vitro maturation culture solution and application thereof
  • Porcine oocyte in-vitro maturation culture solution and application thereof
  • Porcine oocyte in-vitro maturation culture solution and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Preparation of Oocyte Maturation Culture Medium

[0029] Control group culture fluid (the oocyte maturation culture fluid that prior art adopts) formula: take TCM-199 base culture fluid as benchmark, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L L, the volume fraction of serum FBS is 10%, the volume fraction of follicular fluid PFF is 10%, the concentration of cysteine ​​is 0.57mmol / L, the concentration of serum gonadotropin PMSG of pregnant horse is 10IU / mL, human chorion The concentration of gonadotropin HCG was 10IU / mL, and the mass concentration of epidermal growth factor EGF was 10ng / mL.

[0030] The formula of the culture solution of the experimental group: based on the TCM-199 basic culture solution, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L, the volume fraction of serum substitute is 5%, half The concentration of cystine is 0.57mmol / L, ...

Embodiment 2

[0057] 1. Preparation of Oocyte Maturation Culture Medium

[0058] Control group culture fluid (the oocyte maturation culture fluid that prior art adopts) formula: take TCM-199 base culture fluid as benchmark, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L L, the volume fraction of serum FBS is 10%, the volume fraction of follicular fluid PFF is 10%, the concentration of cysteine ​​is 0.57mmol / L, the concentration of serum gonadotropin PMSG of pregnant horse is 10IU / mL, human chorion The concentration of gonadotropin HCG was 10IU / mL, and the mass concentration of epidermal growth factor EGF was 10ng / mL.

[0059] The formula of the culture solution of the experimental group: based on the TCM-199 basic culture solution, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L, the volume fraction of serum substitute is 5%, half The concentration of cystine is 0.57mmol / L, ...

Embodiment 3

[0071] 1. Preparation of Oocyte Maturation Culture Medium

[0072] Control group culture fluid (the oocyte maturation culture fluid that prior art adopts) formula: take TCM-199 base culture fluid as benchmark, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L L, the volume fraction of serum FBS is 10%, the volume fraction of follicular fluid PFF is 10%, the concentration of cysteine ​​is 0.57mmol / L, the concentration of serum gonadotropin PMSG of pregnant horse is 10IU / mL, human chorion The concentration of gonadotropin HCG was 10IU / mL, and the mass concentration of epidermal growth factor EGF was 10ng / mL.

[0073] The formula of the culture solution of the experimental group: based on the TCM-199 basic culture solution, the concentration of sodium bicarbonate is 1.99mmol / L, the concentration of sodium pyruvate is 0.91mmol / L, the volume fraction of serum substitute is 5%, half The concentration of cystine is 0.57mmol / L, ...

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Abstract

The invention discloses a porcine oocyte in-vitro maturation culture solution. The culture solution comprises an insulin transferrin selenium solution ITS, a fibroblast growth factor FGF, a leukemia inhibition factor LIF and inositol. When the culture solution is applied to oocyte in-vitro maturation culture, serum and follicular fluid do not need to be added, and the culture solution is safer, more stable and more efficient.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a porcine oocyte maturation culture solution in vitro and its application. Background technique [0002] With the development of embryo transfer, frozen semen technology, and cloning transgenic technology, the demand for oocytes is increasing. Since the number of oocytes or embryos obtained by pig superovulation technology is extremely limited, it cannot meet the demand for a large number of high-quality oocytes, and the operation is complicated and the cost is high, so in vitro maturation culture is particularly important. [0003] In vitro maturation of porcine oocytes refers to directly collecting discarded ovaries from slaughterhouses, taking out immature oocytes, and simulating the in vivo maturation environment in vitro to achieve the purpose of maturation. In the existing mature culture system, hormones, serum, growth factors or other small molecular substances are g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/25C12N2500/35C12N2501/115C12N2501/235
Inventor 周荣吴珍芳石俊松罗绿花麦然标纪红美余婉娴蔡更元
Owner 广东中芯种业科技有限公司
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