Chicken small yellow follicle outer membrane layer cell separation and culture method

Abstract

The invention discloses a chicken small yellow follicle outer membrane layer cell separation and culture method. The method comprises the following steps: collecting small yellow follicles; acquiring outer membrane layers of the small yellow follicles; cleaning, shearing, digesting and filtering the outer membrane layers; performing primary culture on outer membrane layer cells for 36 to 48 hours; after changing liquid, continuing to perform culturing for 36-48 hours; and finally, performing subculture on the cultured cells for 24-48 hours. The invention provides a pollution-free method for collecting the small yellow follicles, optimizes a method for obtaining outer membrane layers of the small yellow follicles and culturing membrane cells, and further provides a method for subculturing primary membrane cells. By using the method provided by the invention, the membrane cells with relatively high purity can be separated, the consistency in the growth and development period is high, the cell survival rate is high, the success rate of a cell proliferation detection test is high, the result consistency is high, the practicability is strong, the stability is strong, and the feasibility is high. The small yellow follicles collected by the method provided by the invention have a large quantity of cells, and can be continuously supplied to subsequent experiments, so that the poultry purchase cost is reduced, and the experiment efficiency is improved.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a method for the separation, primary culture and subculture of chicken yolk follicle adventitia cells. Background technique [0002] The egg production performance of poultry mainly depends on the development of follicles in the ovaries. High-yield laying hens will have one follicle recruited every day to enter the grade follicle. Once the follicle enters the grade follicle, it will generally not be atresia; the follicle (Small yellow follicle, SYF) with a size of 5-10mm is just the follicle screening. Whether it can enter the key point of priority selection, it can be seen that the development of small yellow follicles is inseparable from the egg production performance of chickens. Unlike mammals, chicken theca cells can not only provide structural support, synthesize androgen, mediate the interaction between oocytes and granulosa cells, but also synthesize estrogen. Theca cell pro...

AI Technical Summary

Problems solved by technology

[0004] For this reason, the present invention provides a kind of chicken small yellow follicle adventitial layer cell separation and culture method, to solve the problem in the prior art that it is difficult to separate pure small yellow follicle adventitial layer cells. Contamination, non-adherence of cells, etc.

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