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Mouse thyroid epithelial cell isolation and culture method

A technology of epithelial cells and culture methods, applied in the field of cell biology, can solve the problems of large differences in survival time and function of thyroid cells, and high cost, and achieve the effects of saving digestion time, reducing dosage, and improving survival rate

Inactive Publication Date: 2017-05-03
JIANGYIN CHI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the primary cultured tissues of the thyroid gland are mostly taken from larger mammals, such as pigs, dogs, sheep, rabbits, etc., and the cost is relatively high, and the survival time and function of the cultured thyroid cells in vitro are also quite different.

Method used

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Embodiment Construction

[0019] 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.

[0020] In the present invention, male mice aged 6-8 weeks are selected to separate thyroid epithelial cells. The specific operation is as follows:

[0021] 1. Take 6-8 week-old male mice, kill them by intraperitoneal injection of pentobarbital sodium, fix the mice, sterilize them with 75% alcohol, take out the mouse thyroid tissue connected to the trachea, and place them in pre-cooled sterile Soak and wash in PBS buffer containing double antibody, and peel off thyroid tissue;

[0022] 2. Quickly remove the capsule, blood cells, connective tissue and other attachments on the thyroid gland under a microscope on ice, and transfer the peeled thyroid gland to a new PBS buffer solution for washing t...

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Abstract

The invention provides a mouse thyroid epithelial cell isolation and culture method. The method includes the steps of: (a) conducting intraperitoneal injection of pentobarbital sodium to kill a mouse, performing alcohol disinfection, than taking out a mouse thyroid tissue connected to a trachea, conducting soaking washing in a precooled sterile double-antibody-containing PBS buffer solution, and stripping the thyroid tissue; (b) carrying out mechanical dissociation of the thyroid tissue, then preheating a mixed enzyme solution and performing shock digestion for 30-40min, using Ficoll to separate thyroid mononuclear cells, and conducting centrifugation to obtain cell precipitate; (c) employing a completely mixed medium to resuspend the cell precipitate, performing inoculation in a treated cell bottle, and carrying out differential adhesion to obtain thyroid epithelial cells; and (d) culturing the purified thyroid epithelial cells in the completely mixed medium. The mouse thyroid epithelial cell isolation and culture method provided by the invention can acquire mouse thyroid epithelial cells with the characteristics of high yield, high purity and long survival time.

Description

technical field [0001] The invention belongs to the field of cell biology, in particular to a method for separating and culturing mouse thyroid epithelial cells. Background technique [0002] The thyroid gland is a very important gland in vertebrates. It belongs to the endocrine organ and is located under the thyroid cartilage in the neck of mammals and on both sides of the trachea. Among them, thyroid epithelial cells (also known as follicular cells or principal cells) are responsible for the production and secretion of thyroid hormones in thyroid cells, including tetraiodothyronine (T4) and triiodothyronine (T3), It has important physiological functions of regulating growth and development, basic metabolism and reproductive activities of animals. Abnormal thyroid function can cause thyroid adenoma, thyroid cancer, primary hyperthyroidism and hypothyroidism and other diseases. The in vitro culture of thyroid cells is an important method for the study of thyroid diseases. ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 张清蒋敏张亚洲齐来俊
Owner JIANGYIN CHI SCI
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