Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Method for separating and culturing primary human esophageal epithelial cells

A technology of epithelial cells and culture methods, applied in the field of cell culture of modern biotechnology, can solve the problems of restricting research work, unstable methods, expensive reagents, etc., and achieve the effect of reliable cell resources, simple and easy-to-master technology, and easier preservation

Inactive Publication Date: 2018-05-11
JIANGYIN CHI SCI
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are few normal esophageal epithelial cells available at home and abroad, and there are very few studies on normal esophageal epithelial cell culture methods in vitro, and most of the methods are either unstable, or the steps are complicated and the reagents used are expensive, which seriously restricts the follow-up research work

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for separating and culturing primary human esophageal epithelial cells
  • Method for separating and culturing primary human esophageal epithelial cells
  • Method for separating and culturing primary human esophageal epithelial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0029] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0030] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0031] The experimental instruments and reagents used in this experiment are as follows:

[0032] A set of surgical instruments, an inverted microscope (XDS-1A, Shanghai), a fluorescence microscope (Leica, the United States), a constant temperature incubator (Heraeus, the United States), a low-temperature centrifuge (TD24B-WS, Shanghai), an ultra-low temperature refrigerator (Zhongke Meiling ), pipette gun (Eppendorf, U.S.), electronic analytical balance (Sartorius, U.S.),...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for separating and culturing primary human esophageal epithelial cells. The method comprises the following steps: (1) aseptically taking out 2 cm of a normal oesophagusexcised in the operation of an esophageal cancer patient; (2) longitudinally cutting open the normal oesophagus along the long axis, flattening the cut normal oesophagus, trimming the flattened normal oesophagus to remove subcutaneous connective tissues and muscles, and repeatedly flushing the trimmed normal oesophagus with a pre-cooled PBS buffer solution containing double antibodies; (3) addinga neutral protease to perform digestion; (4) peeling the epithelial layer and the basal layer; (5) cutting epithelial tissues into pieces, and adding collagenase to digest the epithelial tissue pieces; (6) carrying out digestion termination, filtration, centrifugation and washing, re-suspending the obtained material in a conditioned medium, and inoculating a culture bottle with the obtained solution; (7) placing the inoculation bottle in a 37 DEG C and 5% CO2 incubator, and performing culture; (8) carrying out cell passage and purification; and (9) observing and identifying the morphology ofcells. The method for separating and culturing the human esophageal epithelial cells is simple, stable and efficient, and provides a good experimental material for further researches on esophageal mucosal epithelium differentiation, esophageal pathophysiology, toxicology and the cancerogenesis mechanism.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and specifically relates to a method for separating and culturing human esophageal epithelial cells. Background technique [0002] Esophageal cancer is one of the most common malignant tumors in my country, with obvious regional characteristics. The death of esophageal cancer accounts for 16.05% of malignant tumors, and the cause of death ranks fourth among malignant tumors after gastric cancer, liver cancer and lung cancer. The pathogenesis of esophageal cancer has always been one of the hot issues of research, but it has not been fully elucidated yet. In recent years, the research on the malignant transformation of normal esophageal epithelial cells caused by various physical and chemical factors, and finally the development of esophageal cancer has gradually received attention, but the first condition for studying the proliferation, differentiation and malignant t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0037C12N5/0625C12N2500/84C12N2500/90C12N2501/11C12N2501/33C12N2501/39C12N2509/00C12N2533/54
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products