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Gastroesophageal junction smooth muscle cell tissue explant primary culture and identification method

A technique of smooth muscle cell and tissue block method, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of cumbersome experimental steps, lack of identification of connective tissue cells, etc., and achieve the effect of simple culture method

Active Publication Date: 2018-08-07
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 method for primary culture of smooth muscle cells at the esophagogastric junction by tissue block method

Method used

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  • Gastroesophageal junction smooth muscle cell tissue explant primary culture and identification method
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  • Gastroesophageal junction smooth muscle cell tissue explant primary culture and identification method

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Embodiment 1

[0046] The embodiment of the present invention provides a method for primary culture and identification of smooth muscle cells in the esophagogastric junction by the tissue block method, specifically as follows:

[0047] 1. Obtain muscle strips from esophagogastric junction smooth muscle tissue

[0048] 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...

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Abstract

The invention relates to a gastroesophageal junction smooth muscle cell tissue explant primary culture and identification method. The method includes steps: acquiring gastroesophageal junction smoothmuscle tissue muscle strips; digesting the smooth muscle tissue muscle strips by digestive juice to obtain digested smooth muscle tissue blocks; transferring the digested smooth muscle tissue blocks into a culture container to fix the digested smooth muscle tissue blocks to the bottom of the culture container, keeping a gap of 0.5-1cm between every two adjacent digested smooth muscle tissue blocks, and culturing until adherent cells occupy more than 90% of the bottom of the culture container or the adjacent digested smooth muscle tissue blocks contact through new cells. The invention further provides a smooth muscle cell identification method. The gastroesophageal junction smooth muscle cell tissue explant primary culture method is simple, perfect and effective, and specific phenotypes ofthe smooth muscle cells can be identified by adoption of the identification method.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for primary culture and identification of smooth muscle cells in the esophagogastric junction by tissue block method. 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 esophagogastric 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 food 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 rela...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12Q1/6888G01N33/68
CPCC12N5/0661C12Q1/6888C12Q2600/158G01N33/56966G01N33/68
Inventor 高杨刘俊峰赵连梅张胜雷
Owner FOURTH HOSPITAL OF HEBEI MEDICAL UNIV
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