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Extraction method and application of mouse ovarian tissue fluid

A technology of ovarian tissue and extraction method, which is applied in the field of extraction of mouse ovarian tissue fluid, can solve the problems of low fertilization rate and poor embryonic development ability, and achieve the effect of improving the cleavage rate

Inactive Publication Date: 2011-09-07
上海斯莱克实验动物有限责任公司
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Problems solved by technology

[0005] In order to overcome the problems of low fertilization rate of in vitro mature eggs and poor developmental ability of embryos after fertilization, the present invention provides a method for extracting mouse ovarian tissue fluid and its application

Method used

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  • Extraction method and application of mouse ovarian tissue fluid
  • Extraction method and application of mouse ovarian tissue fluid
  • Extraction method and application of mouse ovarian tissue fluid

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Embodiment 1

[0034] 1. Hormone preparation and storage

[0035] Pregnant horse serum (PMSG) and human chorionic gonadotropin (hCG) were prepared into a solution with a concentration of 50 IU / mL with sterilized physiological saline, and stored at -20°C for future use.

[0036] 2. Preparation of preculture medium

[0037] The mouse ovary is small in size, and the diameter of the ovary of an adult mouse is about 3-4mm, and the tissue fluid content is even less. It is not feasible and expensive to extract pure ovarian tissue fluid. Crush and centrifuge in the culture medium to dissolve the ovarian tissue fluid in the basic culture medium. This extraction method has simple steps and is convenient to operate.

[0038] The specific method is to use 4-week-old female mice that are superovulated in our laboratory for breeding, intraperitoneally inject PMSG 10IU / only, and then intraperitoneally inject hCG10IU / only after 48 hours, separate the mouse ovaries after about 15 hours, and pre-balance was...

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Abstract

The invention provides an extraction method of a mouse ovarian tissue fluid. The method comprises the following steps: placing mouse ovaries in a basal medium; mashing and dissloving the ovarian tissue fluid in the basal medium to form a pre-culture solution. The invention also provides an in-vitro maturation culture method of mouse oocyte, and the method comprises: transferring an immature mouse oocyte into a mature culture solution for in-vitro culture, wherein the mature culture solution is formed by adding the pre-culture solution in the basal medium of oocytes. In addition, the invention also provides a method for obtaining a test-tube mouse through embryo transplantation, and the method comprises: carrying out in-vitro fertilization and embryo transplantation on the oocyte matured by the in-vitro maturation culture method. By using the ovarian tissue fluid provided by the invention in the in-vitro maturation of the mouse oocyte, the cleavage rate of the in-vitro fertilized mature egg can be effectively increased, and is nearly the same as the that of the in-vivo mature egg; and finally, the test-tube mouse can be obtained through embryo transplantation.

Description

technical field [0001] The invention relates to in vitro culture of mouse oocytes, in particular to a method for extracting mouse ovarian tissue fluid and its application. Background technique [0002] Today, with the vigorous development of life science research, the role of experimental animals in scientific research has become increasingly prominent. Mice are one of the most widely used experimental animals. As an inbred mouse, the C57BL / 6J strain mouse is commonly used in reproductive biology, oncology, physiology, immunology and genetics research because of its stable genetic background. Most of the transgenic mice are produced with C57BL / 6J mice as the genetic background. How to scientifically preserve these genetically engineered mice and establish an effective conservation program for mouse germplasm resources is particularly important. At present, the combination of in vitro fertilization technology and embryo freezing technology has been maturely applied to the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075A61D19/00
Inventor 周冬梅刘丽均徐平芮荣郁丽丽朱权凤
Owner 上海斯莱克实验动物有限责任公司
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