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A kind of primary culture and identification method of esophagogastric junction smooth muscle cells by enzymatic digestion

A technology of smooth muscle cells and enzymatic digestion, which is applied in the field of primary culture and identification of smooth muscle cells at the esophagus-gastric junction by enzymatic digestion, can solve the problems of cumbersome experimental steps and lack of identification of connective tissue cells, and achieves the effect of a simple culture method.

Active Publication Date: 2020-08-04
FOURTH HOSPITAL OF HEBEI MEDICAL UNIV
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Problems solved by technology

[0003] Aiming at the technical problems of cumbersome experimental steps and lack of detailed identification of the connective tissue cells involved in the prior art, the present invention provides a primary culture method of smooth muscle cells in the esophagogastric junction by enzymatic digestion

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  • A kind of primary culture and identification method of esophagogastric junction smooth muscle cells by enzymatic digestion
  • A kind of primary culture and identification method of esophagogastric junction smooth muscle cells by enzymatic digestion
  • A kind of primary culture and identification method of esophagogastric junction smooth muscle cells by enzymatic digestion

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Embodiment 1

[0044] The embodiment of the present invention provides a primary culture and identification method of smooth muscle cells in the esophagogastric junction by enzymatic digestion, specifically as follows:

[0045] 1. Obtain muscle strips from esophagogastric junction smooth muscle tissue

[0046] The smooth muscle tissues were obtained from 24 patients with high esophageal cancer. Under the condition of obtaining the approval of the ethics committee and the informed consent of the patient himself or the legally authorized person, the tissue of the esophagogastric junction resected during the operation of the patient is obtained. 15min. Turn the specimen over to the mucosal surface, scrub the entire mucosal tissue with povidone iodine, and then sharply separate the mucosal layer with sterile tissue scissors. According to the course of the muscle fibers, the esophageal circular muscle (EC), esophageal longitudinal muscle (EL), sling fiber (S), hook fiber (clasp fiber, C), proxi...

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Abstract

The invention relates to primary culture and identification methods for smooth muscle cells of esophagogastric junction via enzyme digestion method. The primary culture method includes steps of: 1) acquiring a muscle strip of smooth muscle tissue of the esophagogastric junction; 2) digesting the muscle strip of the smooth muscle tissue with a digestion liquid to prepare a smooth muscle tissue cellsuspending liquid; 3) placing the smooth muscle tissue cell suspending liquid in a culture container, in which levo-polylysine is pre-paved, and culturing the cells until adherent cells account for more than 90% of the bottom of the container or cells grow crowdedly locally. The primary culture method in the invention is simple, completed and effective. The identification method can identify thespecial phenotypes of the smooth muscle cells.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a primary culture and identification method of smooth muscle cells in the esophagogastric junction by enzymatic digestion. Background technique [0002] Primary esophageal motility disorders (PEMD) include achalasia (EAC), diffuse esophageal spasm (DES), nutcracker esophagus (NE), and lower esophageal sphincter hypertension (HLES), all of which occur in the esophagus From the middle and lower segment to the esophageogastric junction (EGJ), the abnormal peristalsis of the esophagus in the diseased segment is characterized by hypertensive peristalsis, and the driving force of the esophagus to the bolus is weakened or disappeared. The clinical manifestations are chronic intermittent chest pain and dysphagia . Because its clinical manifestations are directly related to esophageal motility, it is very important to study the regulation mechanism of smooth muscle tissue relaxation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077C12Q1/6888G01N33/68
CPCC12N5/0661C12N2533/32C12Q1/6888C12Q2600/158G01N33/56966G01N33/68
Inventor 刘俊峰高杨张超刘亮
Owner FOURTH HOSPITAL OF HEBEI MEDICAL UNIV
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