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A kind of porcine oocyte maturation medium in vitro and its preparation method and application

A technology for in vitro maturation and cultivation of oocytes, which is applied in the field of porcine oocyte in vitro maturation culture medium and its preparation, can solve the problems of low in vitro maturation and development rates of porcine oocytes, and achieve improved cleavage rate and improved The effect of mature quality and improved level

Active Publication Date: 2021-03-12
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the current problem of low maturation rate and development rate of pig oocytes in vitro, the invention provides a culture medium for in vitro maturation of pig oocytes and its preparation method and application

Method used

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  • A kind of porcine oocyte maturation medium in vitro and its preparation method and application
  • A kind of porcine oocyte maturation medium in vitro and its preparation method and application
  • A kind of porcine oocyte maturation medium in vitro and its preparation method and application

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Embodiment 1

[0022] Example 1. Oocyte maturation medium in vitro.

[0023] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0024] Based on TCM-199, the mass concentration of penicillin is 0.06g / L, the mass concentration of streptomycin is 0.1g / L, NaHCO 3 The concentration of Hepes is 1.99mmol / L, the concentration of Hepes is 9.8mmol / L, the mass concentration of polyvinyl alcohol is 3g / L, the concentration of sodium pyruvate is 0.91mmol / L, the mass concentration of insulin is 10ng / mL, cysteine The concentration of amino acid is 0.6mmol / L, the mass concentration of epidermal growth factor is 10ng / mL, the volume concentration of porcine follicular fluid is 100mL / L, the concentration of PMSG is 10IU / mL, the concentration of hCG is 10IU / mL, rametide The concentration of amine is 10 -11 mol / L.

[0025] The preparation method of the porcine oocyte maturation medium in vitro described in the present embodiment, concrete step...

Embodiment 2

[0029] Example 2. Oocyte maturation medium in vitro.

[0030] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0031] Based on TCM-199, the mass concentration of penicillin is 0.048g / L, the mass concentration of streptomycin is 0.12g / L, NaHCO 3 The concentration of Hepes is 1.95mmol / L, the concentration of Hepes is 10mmol / L, the mass concentration of polyvinyl alcohol is 2.5g / L, the concentration of sodium pyruvate is 0.95mmol / L, the mass concentration of insulin is 12ng / mL, cysteine The concentration of amino acid is 0.5mmol / L, the mass concentration of epidermal growth factor is 8ng / mL, the volume concentration of porcine follicular fluid is 125mL / L, the concentration of PMSG is 12IU / mL, and the concentration of hCG is 8IU / mL. The concentration of amine is 10 -9 mol / L.

[0032] The preparation method of the porcine oocyte maturation medium in vitro described in the present embodiment, concrete steps are...

Embodiment 3

[0036] Example 3. Oocyte maturation medium in vitro.

[0037] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0038] Based on TCM-199, the mass concentration of penicillin is 0.066g / L, that of streptomycin is 0.08g / L, and that of NaHCO 3 The concentration of Hepes is 2mmol / L, the concentration of Hepes is 9.5mmol / L, the mass concentration of polyvinyl alcohol is 3.5g / L, the concentration of sodium pyruvate is 0.9mmol / L, the mass concentration of insulin is 8ng / mL, cysteine The concentration of amino acid is 0.7mmol / L, the mass concentration of epidermal growth factor is 12ng / mL, the volume concentration of porcine follicular fluid is 75mL / L, the concentration of PMSG is 8IU / mL, and the concentration of hCG is 12IU / mL. The concentration of amine is 10 -7 mol / L.

[0039] The preparation method of the porcine oocyte maturation medium in vitro described in the present embodiment, concrete steps are as follow...

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Abstract

The invention discloses a porcine oocyte in-vitro maturation culture solution as well as a preparation method and application thereof and belongs to the technical field of oocyte in-vitro maturation culture. In order to solve the problems of low porcine oocyte in-vitro maturation rate and development rate at present, the invention provides the porcine oocyte in-vitro maturation culture solution aswell as the preparation method and application thereof. The culture solution comprises a base culture solution TCM-199, penicillin, streptomycin, NaHCO3, 4-hydroxyethylpiperazine ethane sulfonic acid, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, an epidermal growth factor, a porcine follicular fluid, pregnant mare serum gonadotropin, human chorionic gonadotropin and ramelteon. The culture solution can increase the oocyte in-vitro maturation quality, the cumulus cell diffusion index, the glutathione level, the parthenogenetic embryo blastocyst rate and blastocyst total cell number, the in-vitro fertilization embryo cleavage rate, the blastocyst rate and blastocyst total cell number and decrease the ROS level of oocytes.

Description

technical field [0001] The invention belongs to the technical field of in vitro maturation and culture of oocytes. Specifically, it relates to a porcine oocyte maturation culture medium in vitro and a preparation method and application thereof. Background technique [0002] Modern biological and biomedical studies have shown that the biological and genetic characteristics of pigs have a high degree of homology with humans, and are considered to be the preferred animal for the source of human xenogeneic organs. Therefore, pigs are important model animals in biomedical research. The effective combination of in vitro culture technology and today's rapidly developing gene editing technology not only promotes the development and application of embryo engineering, but also drives the development of the medical field. In vitro maturation (IVM) of oocytes is an important part of in vitro embryo production (IVP), and the standards for in vitro maturation (IVM) of porcine oocytes are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/999
Inventor 金君学孙婧陶
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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