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Application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles

A technology for in vitro maturation and culture of oocytes, applied in animal cells, germ cells, vertebrate cells, etc., can solve the problems of maturation rate and mature quality decline, and achieve the effect of increasing the maturation rate

Active Publication Date: 2015-04-01
JILIN UNIV
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Problems solved by technology

Therefore, for large follicle oocyte culture, the inhibition of polyadenylation will lead to the decline of maturation rate and maturation quality.

Method used

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  • Application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles
  • Application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles
  • Application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles

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Embodiment Construction

[0023]

[0024] TCM-199 was purchased from GIBCO; Hepes, PVA, PMSG, EGF, hCG, D-glucose, sodium ketoacid, cysteine ​​and other drugs were purchased from sigma.

[0025] The method for in vitro maturation of pig follicle oocyte of the present invention is as follows:

[0026] (1) Collection of porcine small follicle oocyte cumulus complex

[0027] The ovaries are collected from the ovaries of freshly slaughtered sows, placed in 37°C physiological saline containing 75mg / ml penicillin and 50mg / ml streptomycin, and washed 2-3 times with physiological saline within 2 hours; Use a syringe with a 18-gauge needle indoors to extract the oocyte-cumulus complex with a diameter of <3 mm follicles, and pick out cumulus oocytes (COCs) with 3 to 5 layers of granulosa cells wrapped in a uniform cytoplasm and complete zona pellucida under a microscope Complex;

[0028] (2) Preparation and culture of porcine small follicle oocyte maturation medium in vitro

[0029] Prepare the culture med...

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Abstract

The invention relates to application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles, which belongs to the field of auxology of animals. Degrees of polyadenylation of the maternal genes consisting of cyclinB1, C-mos, GDF9, BMP15 and Cdc2 in oocytes determine expression levels of the genes, which has certain promotion effects on the recruitment process where small follicles become dominant follicles and on maturation of oocytes. The expression levels of the genes consisting of C-mos, GDF9 and BMP15 play a leading role in maturation of the oocytes of small follicles. Addition of a 3'-dA inhibitor can improve the expression levels of the genes consisting of C-mos, GDF9 and BMP15, so the maturation rate of the small follicles is increased. The technology of in-vitro maturation culture of oocytes is expected to be popularized and applied in research fields like transgenic technology, nuclear transplantation technology and embryonic stem cell technology.

Description

technical field [0001] The invention belongs to the field of animal development, and relates to the application of cordycepin in in vitro maturation of porcine follicle oocytes, specifically the GVBD, MI, and MII stages of in vitro maturation of porcine follicle oocytes by cordycepin , inhibiting the adenylation of maternal genes to varying degrees, resulting in differences in the expression of some maternal genes and in vitro maturation rates of porcine small follicle oocytes. Background technique [0002] With the in-depth research on transgenic technology, nuclear transfer technology and embryonic stem cell technology, the rapid development of in vitro fertilization technology has been further promoted. However, the number of oocytes is limited, and only a small number of oocytes are used in the research of embryo transfer technology, which greatly limits the application of new reproductive technologies. The in vitro maturation technology of oocytes can not only reproduc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075C12R1/91
Inventor 高妍刘慧玉张嘉保张永宏翟博张连江袁宝权福实刘殿峰
Owner JILIN UNIV
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