Human primary gastric cancer cell separation and culture method

A technology of gastric cancer cells and culture methods, which is applied in the field of separation and culture of primary human gastric cancer cells, can solve the problems of low cell purity, unfavorable practical application, and long culture time, and achieve high cell purity, good cell growth state, The effect of short digestion time

Inactive Publication Date: 2018-05-25
JIANGYIN CHI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, most of the established methods for the isolation and culture of human gastric cancer cells are based on tissue block culture. The isolated cells are not high in purity, small in number, and have a long culture time.
Regarding the culture of human gastric cancer cells, the methods introduced at home and abroad have problems such as poor reproducibility or complexity, which is not conducive to practical application

Method used

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  • Human primary gastric cancer cell separation and culture method
  • Human primary gastric cancer cell separation and culture method
  • Human primary gastric cancer cell separation and culture method

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Embodiment Construction

[0036] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0037] 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.

[0038] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0039] The experimental instruments and reagents used in this experiment are as follows:

[0040] A set of surgical instruments, inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifuge (TD24B-WS, Shanghai), ultra-low temperature refrigerator (Zhongke Meiling), pipette gun (Eppendorf, USA) ), electronic analytical balance (Sartorius, USA), ultra-clean bench (HJ-CJ-1D, Shanghai), CO 2 Cell culture incubator (SANYO MCO-17AI, Japan)....

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Abstract

The invention provides a human primary gastric cancer cell separation and culture method. The method comprises the following steps of (1) disinfecting a tissue processing device; (2) using a pair of operating scissors for separating a gastric cancer tissue, placing the tissue into a precooled sterile PBS liquid containing double antibodies, and washing for multiple times to remove bloodstains andnecrotic tissues; (3) cutting up the tissues, and using preheated IV type collagenase and trypsin for digesting respectively; (4) terminating digestion, centrifuging, and removing a supernatant; (5) after carrying out cell counting, inoculating into a coated culture flask; (6) culturing at 37 DEG C in an environment with 5 percent of CO2; (7) purifying the cells; (8) subculturing the tissues; (9)observing cell morphology; (10) determining cell survival rate; (11) carrying out immunological identification. The invention provides the human primary gastric cancer cell separation and culture method which is simple to operate, the number of the obtained cells is large, the survival rate is high, the method is the ideal human gastric cancer cell primary separation and culture method, and a reliable cell resource is provided for experiments.

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 primary gastric cancer cells. Background technique [0002] Gastric cancer is a malignant tumor originating from the gastric mucosal epithelium. It ranks first among malignant tumors in China. In my country, the detection rate of early gastric cancer is extremely low, only about 10%. The treatment effect of middle and advanced gastric cancer is poor, and its definite etiology is still uncertain. Prevention and early diagnosis and treatment of gastric cancer are the key to reducing the incidence and death of gastric cancer. [0003] Gastric cancer cell lines include: HGC-27, KATO-III in Europe; AGS, SNU-1, HS764T-1 in the United States; BGC-823, SGC-7901, MGC-803 in China; MKN-1, MKN-28 in Japan , MKN-45, MKN-74, NMGC-3, etc. [0004] At present, cell lines are mostly used in the research of ga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/33C12N2509/00C12N2533/32
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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