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: 2020-10-16
WENS FOOD GRP CO LTD
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  • Abstract
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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 an...

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

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Embodiment Construction

[0024] Below in conjunction with accompanying drawing, invention is described in further detail.

[0025] 1. Oocyte collection and maturation culture

[0026] Obtain the ovaries of young sows from the slaughterhouse, put them in 32°C normal saline, and transport them back to the laboratory. After washing 3 times with physiological saline with added antibiotics, extract 2-6mm follicles and follicular fluid with a 10mL syringe equipped with an 18G needle. Collect in a 50mL conical centrifuge tube, discard the supernatant after natural sedimentation for 10min, resuspend the pellet in DPBS containing 0.05% (wt / vol) PVA, collect more than 3 layers of cumulus cells with uniform cytoplasm under a stereo microscope The cumulus-oocyte complexes (Cumulus oocyte complexes, COCs) were washed three times in the maturation medium, and the COCs were transferred to an incubator containing 500 μL of the maturation medium and kept at 39°C, 5% CO2, and saturated humidity. Equilibrate overnight ...

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Abstract

The invention discloses an oocyte in-vitro maturation culture solution additive. The in-vitro maturation culture solution additive contains ceruloplasmin. Therefore, a first polar body excretion rateof oocytes can be significantly increased, the ROS content of the oocytes can be significantly reduced, and the embryonic blastocyst rate after parthenogenetic activation can be significantly increased, so that the maturation rate and quality of the oocytes can be greatly 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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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/998
Inventor 石俊松吴珍芳周荣麦然标罗绿花蔡更元纪红美余婉娴
Owner WENS FOOD GRP CO LTD
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