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A method for isolation and culture of goose ovarian follicle primary granulosa cells

A technology for granulosa cells and goose ovary, which is applied in the field of separation and culture of primary granulosa cells of goose ovary follicles, can solve the problem of not perfecting the separation and culture method of primary granulosa cells of goose ovary follicles, and achieves low price, simple and easy operation. , the effect of high survival rate

Inactive Publication Date: 2017-03-15
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides a method for isolating and culturing primary granulosa cells of goose ovarian follicles, which solves the problem that there is no perfect method for isolating and culturing primary granulosa cells of goose ovarian follicles

Method used

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  • A method for isolation and culture of goose ovarian follicle primary granulosa cells
  • A method for isolation and culture of goose ovarian follicle primary granulosa cells
  • A method for isolation and culture of goose ovarian follicle primary granulosa cells

Examples

Experimental program
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Effect test

Embodiment 1

[0031] The method for isolating and culturing goose primary granulosa cells comprises the following steps.

[0032] (1) Bloodletting female Sichuan white geese during the egg-laying period was killed, wiped clean with 75% alcohol cotton, cut the abdominal skin to take out the ovaries and follicles, washed away the blood stains with PBS, and put the clean ovarian follicles into In a 500mL beaker filled with PBS, quickly put into the cell culture chamber.

[0033] (2) Take out the larger grade follicles and small yellow follicles, peel off the outer connective tissue and vascular network, wash the blood stains in a 500mL beaker with PBS, and put them in a petri dish with PBS.

[0034] (3) Cut off the stripped follicle in the avascular area opposite the follicle stalk, squeeze out the yolk component with a little force, the granulosa cell layer will be taken out by the yolk and stay on the surface of the yolk, then carefully clip it out with tweezers and put it in In a new Petri...

Embodiment 2

[0046] experimental method.

[0047] (1) Bloodletting female Sichuan white geese during the egg-laying period was killed, wiped clean with 75% alcohol cotton, cut the abdominal skin to take out the ovaries and follicles, washed away the blood stains with PBS, and put the clean ovarian follicles into In a 500mL beaker filled with PBS, quickly put into the cell culture chamber.

[0048] (2) Take out the larger grade follicles and small yellow follicles, peel off the outer connective tissue and vascular network, wash the blood stains in a 500mL beaker with PBS, and put them in a petri dish with PBS.

[0049] (3) Cut off the stripped follicle in the avascular area opposite the follicle stalk, squeeze out the yolk component with a little force, the granulosa cell layer will be taken out by the yolk and stay on the surface of the yolk, then carefully clip it out with tweezers and put it in In a new Petri dish filled with PBS solution.

[0050] (4) The granulosa cells isolated for ...

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Abstract

The invention discloses an isolated culture method of primary granular cells of a goose ovarian follicle. The isolated culture method comprises the following steps: killing a goose by bleeding the carotid artery of the goose, taking out the ovaries of the goose and washing out blood stains by using a phosphate buffer solution; completely removing the connective tissue and vascular net at the outer layer of a follicle; cutting out the follicle in a zone without blood vessels on the opposite side of a follicle handle, slightly pushing to squeeze out yolk and separating granular cells; digesting by using a collagenase in a water bath; adding a fetal calf serum to terminate digestion; filtering to obtain a granular cell suspension; repeatedly centrifuging and washing, collecting cells and depositing the cells in an improved Dubos Eagle culture medium containing 3% of fetal calf serum; inoculating to a cell culture dish, culturing until the cells adhere to the wall of the cell culture dish, and thus obtaining the primary granular cells of the goose ovarian follicle. The invention provides the isolated culture method of the primary granular cells of the goose ovarian follicle, and by adopting the scheme, the isolated culture method is capable of well isolating the granular cells and obtaining relatively pure granular cells, and has the advantage of being practicable, excellent in operability, stable and reliable, good in repeatability and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating and culturing primary granulosa cells of goose ovary follicles. Background technique [0002] In recent years, with the rapid development of the goose industry, the low egg production performance of goose (generally, the annual egg production is only 40-90 eggs) has become an important factor restricting the development of goose industry, and it is one of the key problems that the goose industry needs to solve urgently. one. The egg-laying performance of geese mainly depends on the growth and development level of follicles in the ovary, and is affected by factors such as breed, environment and feeding management. Goose follicles establish a strict hierarchy through development, selection, and atresia. Primordial follicles develop into the small yellow follicle bank through pre-hierarchical development, and then are selected and enter the hierarchal development...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
Inventor 康波马容姜冬梅何珲白林王迅赵玲
Owner SICHUAN AGRI UNIV