Nutrient solution and culturing method for maturation of oocyte in vitro

A technology for in vitro maturation and culture of oocytes, applied in germ cells, animal cells, vertebrate cells, etc., can solve the problem of low developmental ability of oocytes, reduce stress damage, simple culture methods, and improve embryonic development effect of ability

Inactive Publication Date: 2015-09-23
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the problem of low in vitro development ability of existing oocytes, t

Method used

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  • Nutrient solution and culturing method for maturation of oocyte in vitro
  • Nutrient solution and culturing method for maturation of oocyte in vitro

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Experimental program
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specific Embodiment approach 1

[0013] Specific embodiment 1: In this embodiment, the oocyte maturation medium in vitro is composed of conventional oocyte culture medium and vitamin B 12 , fetal bovine serum, FSH, LH, 17β-estradiol, sodium pyruvate, EGF, penicillin and streptomycin sulfate; vitamin B in oocyte maturation medium in vitro 12 The concentration of serum is 2mg / ml, the concentration of serum is 0.1ml / ml, the concentration of follicle stimulating hormone (FSH) is 0.05IU / ml, the concentration of luteinizing hormone (LH) is 0.05IU / ml, the concentration of 17β-estradiol The concentration was 1 μg / ml, the concentration of sodium pyruvate was 24.2 mg / L, the concentration of epidermal growth factor (EGF) was 10 ng / ml, the concentration of penicillin was 100 IU / ml, and the concentration of streptomycin sulfate was 100 μg / ml.

specific Embodiment approach 2

[0014] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the normal oocyte culture medium is DMEM culture fluid, TCM199 culture fluid or MEM culture fluid. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0015] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the serum is calf serum, fetal bovine serum or neonatal bovine serum. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention discloses a nutrient solution for maturation of oocyte in vitro and a method for culturing oocyte using the nutrient solution, and relates to a nutrient solution for nutrient solution for oocyte and a method for culturing the oocyte with the nutrient solution. The nutrient solution and the culturing method using the nutrient solution solve the problem that the viability of oocyte culturing in vitro is low. The nutrient solution for maturation of oocyte in vitro is composed of normal nutrient solution of oocyte, vitamin B12, fetal calf serum, FSH, LH, 17 betas estradiol, sodium pyruvate, EGF, penicillin and streptomycin sulphate. The oocyte is placed into the utrient solution for maturation of oocyte in vitro, the maturation culture on the oocyte lasts for 20-24 hours in the environment in which the volume concentration of CO2 is 5%, and a mature oocyte is acquired. The nutrient solution for maturation of oocyte in vitro can decrease stress injury caused by the external environment, the cytoplasm maturing of the oocyte is promoted, and the parthenogenetic activation of the matured oocyte matured in vitro and the embryos developmental potential after being fertilized are improved.

Description

technical field [0001] The invention relates to an oocyte culture solution and a culture method. Background technique [0002] Obtaining high-quality in vitro mature oocytes is the premise and key to the success of embryo engineering technologies such as animal embryo production in vitro and transgenic cloning. Although the technology of in vitro maturation of oocytes has made great progress in recent years, the development ability of mature oocytes in vitro is still much lower than that of oocytes in vivo. During the process of in vitro maturation of oocytes, affected by the external environment, it is very prone to stress damage, resulting in a decline in developmental ability. [0003] Vitamin B 12 Cobalamin, also known as cobalamin, is the only vitamin containing metal elements, vitamin B 12 It is the latest type of B vitamins discovered so far. B Vitamins in Nature 12 All are synthesized by microorganisms, and higher animals and plants cannot produce vitamin B 12 ...

Claims

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Application Information

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IPC IPC(8): C12N5/075
Inventor 曹新燕许保增李晓霞刁云飞王世勇薛海龙赵伟刚魏海军赵蒙徐超杨镒峰鞠妍邢明杰
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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