Primary culture and identification methods for smooth muscle cells of esophagogastric junction via enzyme digestion method

A technology of smooth muscle cells and enzymatic digestion method, which is applied in the field of primary culture and identification of smooth muscle cells in the esophagogastric junction by enzymatic digestion method, which can solve the problems of cumbersome experimental steps and lack of identification of connective tissue cells, and achieve the effect of simple culture method

Active Publication Date: 2018-06-01
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 i

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  • Primary culture and identification methods for smooth muscle cells of esophagogastric junction via enzyme digestion method
  • Primary culture and identification methods for smooth muscle cells of esophagogastric junction via enzyme digestion method
  • Primary culture and identification methods for smooth muscle cells of esophagogastric junction via enzyme digestion method

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Example Embodiment

[0043] Example 1

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

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

[0046] The smooth muscle tissue came from 24 patients with high esophageal cancer. Subject to the approval of the ethics committee and the patient’s or legally authorized person’s informed consent, samples are taken from the esophagogastric junction tissue excised during the operation of the patient. The proximal blood supply vessel should be disconnected within 40 minutes, and the tissue will not exceed 15min. Turn the specimen over to the mucosal surface, scrub all the mucosal tissue with iodophor, and separate the mucosal layer with sterile tissue scissors. Judging the esophageal circular muscle (EC), esophageal longitudinal muscle (EL), sling fiber (S), c...

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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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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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