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Oocyte in-vitro maturation culture solution and application thereof

An in vitro mature culture and oocyte technology, applied in the direction of cell culture active agent, germ cells, culture process, etc., can solve the problems of unclear composition of follicle fluid, unsatisfactory culture efficiency, etc., and achieve better results

Inactive Publication Date: 2019-12-17
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 that follicular fluid with unclear components needs to be added to the culture medium for in vitro maturation of porcine oocytes and the culture efficiency is not ideal, the present invention provides a culture medium for in vitro maturation of oocytes and its application

Method used

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  • Oocyte in-vitro maturation culture solution and application thereof
  • Oocyte in-vitro maturation culture solution and application thereof
  • Oocyte in-vitro maturation culture solution and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0014] Embodiment 1. Oocyte maturation medium in vitro

[0015] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0016] Based on TCM-199, the volume fraction of KSR is 50mL / L, the mass concentration of penicillin is 0.06g / L, the mass concentration of streptomycin is 0.1g / L, and the mass concentration of polyvinyl alcohol is 3g / L. The mass concentration of insulin is 10ng / mL, the mass concentration of epidermal growth factor is 10ng / mL, the concentration of sodium pyruvate is 0.91mmol / L, the concentration of NaHCO3 is 1.99mmol / L, and the concentration of Hepes is 9.8mmol / L. The concentration of cystine was 0.6mmol / L, the concentration of follicle growth hormone was 10IU / mL, and the concentration of luteinizing hormone was 10IU / mL.

Embodiment 2

[0017] Embodiment 2. Oocyte maturation medium in vitro

[0018] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0019] Based on TCM-199, the volume fraction of KSR is 100mL / L, the mass concentration of penicillin is 0.048g / L, the mass concentration of streptomycin is 0.12g / L, and the mass concentration of polyvinyl alcohol is 2.5g / L , the mass concentration of insulin is 12ng / mL, the mass concentration of epidermal growth factor is 8ng / mL, the concentration of sodium pyruvate is 0.95mmol / L, NaHCO 3 The concentration of Hepes is 1.95mmol / L, the concentration of Hepes is 10mmol / L, the concentration of cysteine ​​is 0.5mmol / L, the concentration of follicle growth hormone is 12IU / mL, and the concentration of luteinizing hormone is 8IU / mL.

Embodiment 3

[0020] Embodiment 3. Oocyte maturation medium in vitro

[0021] The specific components of the oocyte maturation medium in vitro described in the present embodiment are as follows:

[0022] Based on TCM-199, the volume fraction of KSR is 75mL / L, the mass concentration of penicillin is 0.066g / L, the mass concentration of streptomycin is 0.08g / L, and the mass concentration of polyvinyl alcohol is 3.5g / L , the mass concentration of insulin is 8ng / mL, the mass concentration of epidermal growth factor is 12ng / mL, the concentration of sodium pyruvate is 0.9mmol / L, NaHCO 3 The concentration of Hepes is 2mmol / L, the concentration of Hepes is 9.5mmol / L, the concentration of cysteine ​​is 0.7mmol / L, the concentration of follicle growth hormone is 8IU / mL, and the concentration of luteinizing hormone is 12IU / mL.

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Abstract

The invention relates to an oocyte in-vitro maturation culture solution and application thereof, and belongs to the technical field of oocyte in-vitro maturation culture solutions. The oocyte in-vitromaturation culture solution and application thereof are provided for solving the problems that it is necessary to add follicular fluid with unclear compositions into the oocyte in-vitro maturation culture solution, and the culture efficiency is not ideal. The culture solution is composed of TCM-199, KSR, penicillin, streptomycin, NaHCO3, Hepes, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, epidermal growth factors, follicle-generating hormones and luteinizing hormones. According to the oocyte in-vitro maturation culture solution, during oocyte in-vitro culture, the similar effects onthe cleavage rate and the blastocyst formation rate can be obtained as a traditional culture solution with added follicular fluid, the disadvantages that in traditional porcine oocyte in-vitro culture, follicular fluid with unknown components needs to be added can be avoided, and thus an oocyte in-vitro maturation system is more standardized.

Description

technical field [0001] The invention belongs to the technical field of oocyte culture. Specifically relates to an oocyte maturation culture medium in vitro 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 production 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 constantly improving. Although the pig oocyte IVM system ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/32C12N2501/11C12N2501/31C12N2501/33C12N2501/999
Inventor 金君学孙婧陶郝耘初张文涵朱彦龙王张钰
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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