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Method for improving in vitro fertilizable competence of vitrification freezing oocyte

A technology of vitrification and oocytes, which is applied in the field of artificial in vitro fertilization, can solve the problems of decreased fertilization ability of vitrified oocytes, solve the problem of decreased fertilization ability, increase cleavage rate and blastocyst rate, and be widely used foreground effect

Active Publication Date: 2018-09-28
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problem of decreased fertilization ability of vitrified oocytes in the prior art, an object of the present invention is to provide the application of gadolinium and CLC in improving the in vitro fertilization ability of vitrified oocytes

Method used

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  • Method for improving in vitro fertilizable competence of vitrification freezing oocyte
  • Method for improving in vitro fertilizable competence of vitrification freezing oocyte
  • Method for improving in vitro fertilizable competence of vitrification freezing oocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Effect of Gadolinium Treatment on Vitrified Bovine Oocyte Cytoplasmic Calcium, Mitochondrial Calcium, Endoplasmic Reticulum Calcium, and CG Distribution

[0061] Bovine oocytes were treated with 50 μM, 150 μM, and 300 μM gadolinium for 40 minutes, and then vitrified. The effects of different gadolinium treatments on the distribution of oocyte cytoplasmic calcium, mitochondrial calcium, endoplasmic reticulum calcium, and CG were compared.

[0062] As shown in Table 1, the cytoplasmic calcium ion concentration and mitochondrial calcium ion concentration in the gadolinium treatment group were significantly lower than those in the vitrification group, the endoplasmic reticulum calcium ion concentration in the gadolinium treatment group was significantly higher than that in the vitrification group, and the cytoplasmic calcium ion concentration in the 150 μM gadolinium treatment group There was no significant difference in internal calcium ion concentration, mitochon...

Embodiment 2

[0067] Example 2 Effect of Gadolinium Treatment on Fertilization Ability of Vitrified Bovine Oocytes

[0068] Bovine oocytes were treated with 50 μM, 150 μM, and 300 μM gadolinium for 40 min, and then vitrified. After thawing, the surviving oocytes were selected for IVF, and the oocytes were tested for fertilization ability after 8 hours of IVF. Then compare the effects of different gadolinium treatments on the fertilization ability of bovine oocytes.

[0069] As shown in Table 2, the normal fertilization rate in the 150 μM gadolinium treatment group (64.29±6.28%) was significantly higher than that in the vitrification group (47.22±4.39%), but still lower than that in the fresh group (78.13±6.72%). Meanwhile, the infertilization rate (26.19±2.41%) in the 150 μM gadolinium treatment group was significantly lower than that in the vitrification group (38.89±3.71%), but still significantly higher than that in the fresh group (15.63±1.30%). The fertilization rate of polyspermia i...

Embodiment 3

[0072] Example 3 Effect of Gadolinium Treatment on Fertilization Efficiency of Vitrified Bovine Oocytes

[0073] Bovine oocytes were treated with 50 μM, 150 μM, and 300 μM gadolinium for 40 min, and then vitrified. After thawing, the surviving oocytes were selected for IVF, and the cleavage rate and blastocyst rate were counted, and then the effects of different gadolinium treatments on the fertilization efficiency of bovine oocytes were compared.

[0074] As shown in Table 3, the cleavage rate and blastocyst rate (62.20±5.63%, 23.53±2.31%) of the 150 μM gadolinium treatment group were significantly higher than those of the vitrification group (40.85±3.82%, 13.79±1.05%), but both were significantly Lower than fresh group (79.03±7.62%, 36.73±3.28%).

[0075] Table 3 Effect of gadolinium on in vitro fertilization efficiency of vitrified bovine oocytes

[0076]

[0077]

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Abstract

The invention provides a method for improving in vitro fertilizable competence of vitrification freezing oocyte. The method disclosed by the invention comprises the following steps: performing combined treatment on bovine oocyte with a solution containing gadolinium and CLC before vitrification freezing treatment, and performing vitrification freezing; treating with methyl-beta-cyclodextrin afterunfreezing, and performing in vitro fertilization. Results prove that the method is capable of obviously improving the cleavage rate and blastocyst rate of vitrification freezing bovine oocyte and achieving in vitro fertilization efficiency higher than that of fresh bovine oocyte, has a wide application prospect and is worthy of popularization.

Description

technical field [0001] The invention relates to the field of artificial in vitro fertilization, in particular to a method for improving the in vitro fertilization ability of vitrified frozen oocytes. Background technique [0002] Oocyte vitrification technology is an important part of livestock embryo biotechnology, which removes time and space constraints for in vitro embryo production, and greatly promotes the efficient use of excellent female genetic resources across time and space. For rare and endangered animal genetic resources Long-term preservation is also of great significance (Zhu Shien, Zeng Shenming, Wu Tongyi, Meng Qinggang, Zhang Zhongcheng, Chen Yongfu. Research on vitrification of bovine oocytes by OPS method. Chinese Agricultural Sciences, 2002; 35(6):700-704. ). At the same time, this technology provides an important methodological guarantee for female fertility preservation (Azari M, Kafi M, Ebrahimi B, FatehiR, Jamalzadeh M. Oocyte maturation, embryo dev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075A01N1/02
CPCA01N1/0221A01N1/0226C12N5/0609C12N2500/20C12N2500/34C12N2500/36C12N2501/392C12N2501/31
Inventor 赵学明朱化彬郝海生赵亚涵杜卫华刘岩庞云渭
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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