Method for separating and culturing mouse ovarian granule cells

A technology of granulosa cells and culture methods, which is applied in the field of separation and culture of mouse ovary granulosa cells, can solve the problems of less research on the in vitro culture of mouse ovary granulosa cells, and achieve the effect of reducing costs and increasing the survival period

Inactive Publication Date: 2017-12-15
JIANGYIN CHI SCI
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  • Claims
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Problems solved by technology

At present, there are different methods for isolating rat ovarian granulosa cells in vitro, but there are few studies on the in vitro culture of mouse ov

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  • Method for separating and culturing mouse ovarian granule cells
  • Method for separating and culturing mouse ovarian granule cells

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Embodiment Construction

[0017] In order to present the purpose and advantages of the present invention more clearly, specific embodiments will now be further described. The specific embodiments described here are only for explaining the present invention, and are not intended to limit the present invention.

[0018] The present invention selects 10-15 g of immature female mice and separates ovarian granulosa cells. The specific operation is as follows:

[0019] 1. Take immature female mice, inject PMSG 5-10IU subcutaneously first, and then inject HCG 5-10IU 40-48h later;

[0020] 2. After 15-20 hours, the mice were killed by neck dislocation, and the mice were fixed. After being disinfected with 75% alcohol, the ovaries were quickly removed aseptically and placed in preheated PBS. For attachments such as tissues, transfer the stripped ovaries to new PBS buffer and wash them twice;

[0021] 3. Transfer the ovary to the mixed basal medium and incubate at 37°C for 10-15min;

[0022] 4. Puncture the ...

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Abstract

The invention provides a method for separating and culturing mouse ovarian granule cells. The method comprises (a) intraperitoneally injecting pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) into a female mouse, (b) killing the mouse through a cervical dislocation method, separating mouse ovarian, carrying out PBS cleaning and removing fat and tunica around the mouse ovarian through a stereoscopic microscope, (c) puncturing follicles in a mixed base culture medium through a syringe needle to release granule cells and oocytes, adding a mixed enzyme into the culture medium, carrying out digestion, carrying out filtration through a screen, centrifuging the filtrate and removing the supernatant and (d) carrying out inoculation with completely mixed medium resuspended cell precipitates. The method realizes simple, efficient and stable separation and culture of ovarian granule cells.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for separating and culturing mouse ovarian granulosa cells. Background technique [0002] Ovarian granulosa cells are a very important type of cells in the reproductive process of animals. As the largest cell group in follicles, they not only cooperate with theca cells to regulate the synthesis of female steroid hormones, but also are closely related to the growth and maturation of oocytes. Therefore, granulosa cells can be used as a good cell model to study the biological behavior and functional regulation of germ cells in females and female animals, as well as to evaluate in vitro drug model systems and regulate the genetic functions of specific genes. At present, there are different methods for isolating rat ovarian granulosa cells in vitro, but there are few studies on the in vitro culture of mouse ovarian granulosa cells. The present invention aims to provide...

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2509/00
Inventor 张清张亚洲蒋敏齐来俊
Owner JIANGYIN CHI SCI
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