Method for separating and culturing pituitary gland cells of rat

Abstract

The invention provides a method for separating and culturing pituitary gland cells of a rat. The method comprises steps as follows: (1), an 8d male SD rat is selected and subjected to anesthesia execution; (2), the anesthetized rat is fixed on a board with the ventral side facing upwards, the pituitary gland of the rat is taken out under the sterile condition, put in a pre-cooled PBS (poly(butylene succinate)) liquid and washed repeatedly, bloodstain is removed, and pituitary gland envelope is removed; (3), the pituitary gland is cut into 1 mm*1 mm fragments by ophthalmic scissors in the pre-cooled PBS liquid under the sterile condition; (4), pre-heated III type collagenase is combined with neutral enzyme for mixed digestion; (5), the digestion is stopped, filtering is performed, a filtrate is collected and centrifuged, a supernatant is abandoned, a culture medium is added for resuspension, and the operation is repeated three times; (6), a 24-hole culture plate is inoculated with cells for culture; (7), the cells cultured for 7d are subjected to trypsin digestion and then are purified by a screen again, a supernatant is abandoned after centrifugation, the cells are resuspended, and the treated culture plate is inoculated with the cells; (8), the cells are subjected to cell immunofluorescence identification. The method for separating and culturing the pituitary gland cells of the rat is simple to operate, the obtained pituitary gland cells are high in survival rate and good in activity, can be used for establishing a cell experimental model in vitro and meet the requirement of pituitary gland cell experiments.

Description

technical field [0001] The invention belongs to the field of cell technology, in particular to a method for separating and culturing rat pituitary cells. Background technique [0002] The pituitary gland, also known as the hypocerebral gland, is an important part of the endocrine system, located on the ventral surface of the diencephalon, behind the optic chiasm, and connected to the hypothalamus. The pituitary gland is the most important endocrine gland in the organism. The anterior pituitary gland is composed of thyrotropin cells, gonadotroph cells, growth hormone cells and other cells. The pituitary gland is dually regulated by the hypothalamus and target organ hormones, and it plays an important role in the metabolism of the body, maintaining the stability of the internal environment, and the growth and development of the body. Pituitary cells cultured in vitro can be used to study the mechanism of stimulating luteinizing hormone (LH) synthesis and secretion, and the r...

AI Technical Summary

Problems solved by technology

At present, there are few studies on the isolation and culture of rat pituitary cells at home and abroad, the isolation method is immature, and the obtained cells are of low purity and small number.

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