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Application of enhancing antioxidation of oocyte cultured in vitro by Peroxiredoxin 4 protein

An oocyte, in vitro culture technology, applied in animal cells, tissue culture, germ cells, etc., can solve problems such as unfavorable oocyte development, improve oocyte fertilization rate and clinical pregnancy rate, delay aging, improve effect of maturity

Pending Publication Date: 2019-03-26
JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no method to improve in vitro maturation of immature eggs by improving oxidative stress in cumulus cell-oocyte complex (COC) granulosa cells
[0005] It is generally believed that the reactive oxygen species (ROS) and oxygen free radicals produced in the follicle, such as OH, O 2 and H 2 o 2 , especially the active oxygen produced during ovulation, which is not conducive to the development of oocytes

Method used

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  • Application of enhancing antioxidation of oocyte cultured in vitro by Peroxiredoxin 4 protein
  • Application of enhancing antioxidation of oocyte cultured in vitro by Peroxiredoxin 4 protein
  • Application of enhancing antioxidation of oocyte cultured in vitro by Peroxiredoxin 4 protein

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Embodiment 1

[0036] The invention provides the use of a Peroxiredoxin4 protein in regulating the balance between reactive oxygen species and anti-oxidation in cultured oocytes in vitro. Further, the use of a peroxiredoxin4 protein in an in vitro culture reagent that plays a regulatory role in resisting cell apoptosis caused by oxidative stress in granulosa cells is proposed. Further, a Peroxiredoxin4 protein is proposed to inhibit H 2 O 2 Use of in vitro culture reagents resulting in decreased cumulus expansion index and oocyte MII maturation rate.

[0037] The in vitro culture reagent (liquid) is an in vitro oocyte culture solution and an in vitro culture and maturation (IVM) culture solution of immature eggs. In addition to the Peroxiredoxin 4 protein, it also includes CZB culture solution, KSOM culture solution, and HTF culture solution. Any kind of culture medium is used as the basal medium. The above-mentioned basal media are all commercial culture reagents known to those skilled i...

Embodiment 2

[0056] Embodiment 2: In order to better understand the present invention, the inventors describe the specific experimental process and experimental data as follows:

[0057] 1. Main instruments and equipment used

[0058] Pipette: Eppendorf;

[0059] CO2 incubator: Forma 521;

[0060] Inverted phase contrast microscope: Nikon 1000;

[0061] Ultra-clean workbench: Su Jing;

[0062] Stereo microscope: Nikon 2000;

[0063] Heating platform: domestic;

[0064] Petri dish;

[0065] 1. Main reagents

[0066] Peroxiredoxin 4 protein: purchased from Abcam Company.

[0067] No Peroxiredoxin 4 protein was added to the IVM solution of group 1;

[0068] Group 2 added Peroxiredoxin 4 protein 20ug / ml to IVM solution;

[0069] Ovarian tissue senescence was artificially induced by irradiating normal female ICR mice with 4 Gy of ionizing radiation.

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Abstract

The invention provides an application of enhancing antioxidation of an oocyte cultured in vitro by a Peroxiredoxin 4 protein. The maturing rate of the immature oocyte cultured in vitro can be improvedby improving the resistance of particle cells in oxidative stress and improving the antioxidation of the oocyte. Aging of the oocyte is also delayed through an antioxidant stress mechanism in the cell, and the fertility rate and the clinical pregnancy rate of the oocyte in an assisted breeding process are improved.

Description

technical field [0001] The invention belongs to the fields of assisted reproduction and embryo culture. Specifically, it relates to the use of Peroxiredoxin 4 protein to enhance the antioxidant effect of cultured oocytes in vitro. Background technique [0002] Oxidative stress / ROS damage caused by aging and changes in the internal environment under pathological conditions will affect the development of follicles. This is because in the pathological state, the oxidative stress of granulosa cells can lead to cell apoptosis, affecting the protective and regulatory functions of granulosa cells, thereby affecting the development of eggs. On the other hand, the mature eggs in vivo or in vitro in assisted reproductive therapy are aged due to oxidative stress in the process of in vitro culture before insemination, which affects the quality of oocytes, fertilization ability and embryonic developmental potential. Reduce clinical pregnancy rates. [0003] In recent years, embryo tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/998
Inventor 孟艳刘嘉茵钱易崔毓桂严正杰
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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