Optimization liquid for in vitro maturation of advanced-age mouse oocytes

An oocyte and in vitro maturation technology, applied in the field of animal reproduction, can solve the problems of reducing the quality of oocytes, achieve simple and clear operation, reduce oocyte degeneration, and improve the effect of in vitro maturation

Pending Publication Date: 2020-09-18
YANGZHOU UNIV
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Problems solved by technology

The reason may be that with age, changes in hormone levels in the body affect the reproductive function of the ovary and reduce the quality of oocytes, s

Method used

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  • Optimization liquid for in vitro maturation of advanced-age mouse oocytes
  • Optimization liquid for in vitro maturation of advanced-age mouse oocytes
  • Optimization liquid for in vitro maturation of advanced-age mouse oocytes

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Experimental program
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Embodiment Construction

[0025] Materials and Methods;

[0026] 1. Test animals and feeding management;

[0027] ICR female mice (10-12 months old) and male mice (8-12 weeks old) were purchased from the Comparative Medicine Center of Yangzhou University, raised in a temperature-controlled and light-controlled environment at 20-25°C, with 12h light and dark intervals, and fed ad libitum With drinking water, after 3-5 days of adaptation, the experiment was carried out, and the operation was in compliance with the relevant regulations on the welfare of experimental animals.

[0028] 2. Main reagents and instruments;

[0029] Pregnant horse serum gonadotropin PMSG (Ningbo Second Hormone Factory), M2 culture medium, paraffin oil, hyaluronidase, melatonin (US Sigma Company); IVM (in vitro maturation medium), TYH (sperm capacitation solution ), HTF (fertilized fluid), KSOM (embryo culture medium) (Nanjing Aibei Biotechnology Company); reactive oxygen species detection kit, Mito-Tracker Red CMXRos mitochondri...

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Abstract

The invention relates to optimization liquid for in vitro maturation of advanced-age mouse oocytes. A preparation method comprises the steps of firstly, weighing 0.0232g of melatonin, enabling the melatonin to dissolve in 1ml of anhydrous ethanol, preparing 100mM of concentrated storage liquid, diluting 100mM of the concentrated storage liquid in an IVM culture medium until the concentration is 10-6mol/L, and the final concentration of the anhydrous ethanol is 0.01% smaller than that of the IVM culture medium, and preparing the optimization liquid for in vitro maturation of advanced-age mouseoocytes. In accordance with in vitro maturation culture liquid for commercial mouse oocytes, the culture efficiency cannot meet production and embryo transplanting requirements, and the in vitro maturation culture liquid for commercial mouse oocytes is improved, so that the maturing rate ( 87.53 +/-2.88% ) and the cleavage rate ( 79.04 +/-3.13% ) after fertilization, and the blastula rate( 65.69 +/-2.07% ) of the advanced-age mouse oocytes can be notably increased. Besides, the distribution and the activity of mitochondrial can be improved, and the oxidation stress level and the early apoptosis are reduced to weaken oocyte degradation caused by aging.

Description

technical field [0001] The invention relates to an optimized solution for in vitro maturation of aged mouse oocytes, which belongs to the field of animal reproduction. Background technique [0002] There are few reports on the in vitro maturation culture of aged mouse oocytes. At present, the commercialized mouse oocyte maturation medium in vitro is directly used in the in vitro culture of aged mice, and the maturation rate can only reach 77.64±2.13 %, the cleavage rate after in vitro fertilization was 41.77±0.59%, and the blastocyst rate was 35.55±2.22%, which were significantly lower than the culture effect of young mice. The reason may be that with age, changes in hormone levels in the body affect the reproductive function of the ovary and reduce the quality of oocytes, so it is necessary to find a culture medium that can optimize the in vitro maturation of commercial mice , to improve the quality of oocytes in aged mice. Contents of the invention [0003] The object ...

Claims

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Application Information

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IPC IPC(8): C12N5/075G01N21/64
CPCC12N5/0609C12N2501/30G01N21/6428
Inventor 李拥军胡慧茹王健马金亮张柳明冯云奎
Owner YANGZHOU UNIV
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