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In vitro maturation culture method of denuded oocytes of mice

An in vitro mature culture and naked egg technology, which is applied in the field of in vitro mature culture of mouse naked eggs, can solve the problems of the blastocyst stage that cannot be further developed to reach the uterine implantation, oocytes do not have the ability to combine with sperm and fertilize, and achieve The effect of promoting application and promotion, high efficiency of maturation and development, and high quality of embryonic development

Inactive Publication Date: 2014-07-02
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previously, in the in vitro maturation of naked eggs in the world, the conventional culture system can lead to the maturation and development of oocytes, but mature oocytes do not have the ability to fertilize with sperm, and cannot further develop in vitro to achieve uterine implantation blastocyst stage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0017] 1 Experimental method

[0018] 1.1 In vitro maturation and culture of naked eggs

[0019] Take the mouse ovary and use the head of a syringe needle under a microscope to puncture the follicle on the ovary, and release the oocytes in the follicle into the TCM199 oocyte in vitro operating solution containing HEPES and 10% FBS. TCM199 is a product of Therma , Product No. SH30233; After washing the naked eggs in vitro with an egg picking needle, randomly put them into oocyte maturation culture medium containing 50μM forskolin In In-Vitro MaturationMedia, naked eggs were cultured in oocyte maturation medium containing 50μM forskolin for 3h, then naked eggs were incubated in oocyte maturation medium containing 2.5μM PD166285 After washing in In-Vitro Maturation Media, transfer to oocyte maturation medium containing 2.5μM PD166285 In-Vitro Maturation Media continued in vitro culture for 16h. among them In-Vitro Maturation Media is a product of Sage in the United States, and the...

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Abstract

The invention discloses an in vitro maturation culture method of denuded oocytes of mice. The method comprises the following steps: (1) performing in vitro maturation culture on the denuded oocytes; (2) performing parthenogenetic activation on matured denuded oocytes; (3) performing in vitro fertilization on the matured denuded oocytes. According to the method, an environment for artificially simulating natural maturation of oocytes can be created, so that the denuded oocytes can efficiently maturate and grow in vitro and can be subjected to in vitro fertilization and embryonic development as same as normally growing oocytes after in vitro maturation culture. The practice proves that the in vitro maturation culture efficiency of the denuded oocytes of the system is high, the system is mature and stable, and the embryonic development quality of the denuded oocytes is high after in vitro natural fertilization, so that the application and popularization of the denuded oocytes in reproductive health of human beings and genetic breeding of modern livestock can be greatly facilitated. In addition, the operation of obtaining blastulae from the denuded oocytes is simplified and the denuded oocytes serving as ovum sources can be conveniently put into actual production.

Description

Technical field [0001] The invention relates to an in vitro maturation culture method of mouse naked eggs. Background technique [0002] In normal reproductive development, a large number of cumulus cells are wrapped around the oocyte. During natural ovulation, the oocytes and the surrounding cumulus cells are discharged into the fallopian tube to wait for the combination with sperm. In embryo engineering operations or assisted reproductive technology applications, a large number of cumulus cells are wrapped around the oocytes with high maturation efficiency, and the mature oocytes have the function of naturally combining with sperm to form an embryo. However, in the oocytes directly extracted from the ovary in clinical applications, more than half of the oocytes are not completely surrounded by cumulus cells. This type of oocytes has low maturation efficiency, and even if they can mature, they will not have it after maturation. The ability to combine with sperm is often directl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
Inventor 卞雅妮曹鸿国章孝荣张飞魏超李运生刘亚张运海方富贵
Owner ANHUI AGRICULTURAL UNIVERSITY
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