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Oocyte in-vitro maturation culture solution additive and application thereof

An in vitro maturation culture and oocyte technology, applied in the field of oocyte in vitro maturation medium additives, can solve the problems of affecting oocyte maturation and development ability, impairing cell survival, accumulation of ROS, etc. Effects of one-polar body excretion rate, increasing blastocyst rate and reducing ROS content

Active Publication Date: 2021-07-23
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in vitro culture is a static environment without the replacement of nutrients and metabolites, which tends to cause the accumulation of ROS, which alters their function and impairs cell survival.
Thus, ROS negatively impact oocyte viability, gene expression, protein synthesis, and molecular signaling, affecting oocyte maturation and developmental competence

Method used

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

Examples

Experimental program
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Effect test

Embodiment 1

[0023] 1. Oocyte collection and maturation culture

[0024] Obtain sheep ovaries from the slaughterhouse, put them in 34°C normal saline, and transport them back to the laboratory. After washing 3 times with normal saline supplemented with 200IU penicillin and streptomycin, extract 2-8mm follicles with a 10mL syringe equipped with an 18G needle. The liquid was collected in a 50mL conical centrifuge tube, and the supernatant was discarded after natural precipitation for 10min, and the precipitate was resuspended in DPBS (Dulbecco's Phosphate-Buffered Saline, Dulbecco's Phosphate-Buffered Saline) containing 0.1% PVA (Polyvinylalcohol, polyvinyl alcohol). Collect cumulus-oocyte complexes (Cumulus oocyte complexes, COCs) with more than 3 layers of cumulus cells and uniform cytoplasm under a stereo microscope. After washing three times in the maturation medium, transfer the COCs to a medium containing 500 μL of the following different The grouped mature culture medium has been incu...

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Abstract

The invention discloses an oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the technical field of biology. Lactoferrin is found to be capable of remarkably improving the first polar body discharge rate of oocyte, remarkably reducing the ROS content of the oocyte and remarkably improving the blastocyst rate of an embryo after in-vitro fertilization, so that the maturation rate and quality of the oocyte can be remarkably improved, and the lactoferrin can be used for preparing the oocyte in-vitro maturation culture solution additive or an oocyte in-vitro maturation culture solution. The additive and the application can be applied to the embryo engineering technology, and the success rate and quality of embryo engineering are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an oocyte maturation culture medium additive in vitro and its application. Background technique [0002] Mammalian in vitro fertilized embryos, cloned embryos, parthenogenetic embryos and transgenic embryos are widely used in the field of embryo engineering technology such as genetic improvement, early embryo development mechanism research, epigenetic regulation, etc. The quality of oocyte culture in vitro is the key to determining embryo development However, so far the maturation efficiency and quality of oocytes cultured in vitro have not been greatly improved. Reactive oxygen species (reactive oxygen species, ROS) is an important factor affecting oocyte culture in vitro. ROS is a product of biological aerobic metabolism, including oxygen ions, superoxide ions, hydroxyl radicals, and hydrogen peroxide. In equilibrium, ROS, as signaling molecules, play beneficial roles in physiolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/24C12N2500/32C12N2501/999C12N2500/30C12N2501/392C12N2501/11C12N2501/31
Inventor 戴雁峰任敬宇郝玉春刘展鹏
Owner INNER MONGOLIA UNIVERSITY
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