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Culture medium for keeping primary airway epithelial cells in physiological state in vivo

A physiological state, epithelial cell technology, applied in the direction of animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of difficult primary cell culture, and achieve the effects of easy promotion and application, wide promotion value, and simple preparation

Inactive Publication Date: 2012-06-20
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitation of this special growth condition, the existing conventional medium cannot create a suitable microenvironment for the growth of respiratory epithelial cells, so it is difficult to culture this type of cells in primary culture.

Method used

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  • Culture medium for keeping primary airway epithelial cells in physiological state in vivo
  • Culture medium for keeping primary airway epithelial cells in physiological state in vivo
  • Culture medium for keeping primary airway epithelial cells in physiological state in vivo

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Experimental program
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Embodiment Construction

[0014] 1. Preparation medium

[0015] In order to facilitate storage and experimental operation, the chemical reagents purchased in this experiment were first prepared into standard storage concentrations and stored under suitable temperature conditions for a long time. The dosage is calculated according to the prepared standard storage concentration. Concrete preparation process is as follows:

[0016] Take penicillin, streptomycin, insulin, transferrin, epidermal growth factor, cholera toxin, hydrocortisone, bovine pituitary extract, bovine serum albumin, retinoic acid, Add amphotericin, amphotericin B and fetal bovine serum to the DMEM / F12 medium, add water to 100ml and mix thoroughly, filter through a 0.22 μm microporous membrane in a laminar flow cell culture chamber, and collect the filtrate to obtain 100ml of the present invention. culture medium.

[0017] Table 1

[0018]

[0019] 2. Culture primary airway epithelial cells

[0020] 1. Petri dish collagen coatin...

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Abstract

The invention relates to preservation of human local living parts and in particular relates to a culture medium for keeping primary airway epithelial cells in the physiological state in vivo during in vitro culture. The invention is characterized in that the culture medium contains the following components (per liter), 12 g of DMEM / F12 medium, 30 mg of penicillin G sodium salt, 50 mg of streptomycin, 10 mg of insulin, 5 mg of transferring, 25 Mug of epidermal growth factors, 100 Mug of cholera toxin, 108.741 Mug of hydrocortisone, 1 g of bovine pituitary extract, 3 g of bovine serum albumin, 1.5*10<-2> Mug of retinoic acid, 50 mg of gentamycin, 0.5 mg of amphotericin B, 50 mL of fetal calf serum and the balance of water. The primary airway epithelial cells cultured by the culture medium have the ciliary beating function and can maintain the physiological state in vivo.

Description

technical field [0001] The present invention relates to the preservation of local living parts of humans, in particular to a culture medium for primary airway epithelial cells. Background technique [0002] Respiratory epithelial cells constitute the first natural barrier from the nasal cavity to the terminal bronchioles, and are the main starting site for inflammatory responses after pathogens invade the body through the respiratory tract. At home and abroad, cell lines such as A549, 16HBE, and BEAS-2B are mostly used to replicate cell models of respiratory diseases to study microbial infections and inflammatory responses. However, cell lines such as A549 and 16HBE are derived from tumor cells, or immortalized cell lines such as BEAS-2B are obtained from the fusion of normal airway epithelial cells and tumor cells, and their physiological characteristics are quite different from those of somatic cells. , the cytological study of respiratory diseases has certain limitations...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 王达李煜生
Owner SOUTHERN MEDICAL UNIVERSITY
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