Method for culturing gastric cancer solid tumor primary cells

A primary cell and cell culture technology, applied in the direction of cell culture active agent, tumor/cancer cell, culture process, etc., can solve the problems of long culture cycle, difficult to remove miscellaneous cells, low culture success rate, etc.

Inactive Publication Date: 2020-10-23
SUZHOU GENOARRAY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing primary tumor cell culture technologies mainly include 2D culture, 3D culture, reprogramming culture, etc. These methods face the problems of extremely long culture cycle, low culture success rate, and difficulty in removing stray cells to varying degrees.

Method used

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  • Method for culturing gastric cancer solid tumor primary cells
  • Method for culturing gastric cancer solid tumor primary cells
  • Method for culturing gastric cancer solid tumor primary cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1. Preparation of reagents for culturing gastric cancer primary cells

[0077] 1. Sample preservation solution (100mL)

[0078] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0079] Table 1 Sample Preservation Solution (100mL)

[0080]

[0081] After the sample preservation solution is prepared, it is divided into 15mL centrifuge tubes, 5mL per tube. After aliquoting, it can be stored at 4°C for 1 month.

[0082] 2. Sample cleaning solution (100mL)

[0083] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0084] Table 2 Sample cleaning solution (100mL)

[0085]

[0086] The sample cleaning solution should be prepared and used immediately.

[0087] 3. Sample dissociation solution (10mL)

[0088] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0089] Table 3 Sample Dissociation Solution (10mL)

[0090]

[0091]

[0092] Note...

Embodiment 2

[0176] Example 2, acquisition of postoperative specimens of gastric cancer

[0177] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0178] 2. The attending doctor selects the patients according to the clinical indications stipulated in the medical guidelines, and selects appropriate samples for in vitro culture according to the clinical indications during the operation. The selection criteria of the samples are: primary gastric cancer, and the pathological stage is stage II , Stage III or IV, various pathological types of gastric cancer or metastatic lesions of gastric cancer, gastric cancer surgical specimens weighing more than 20mg.

[0179] 3. The attending doctor provides basic clinical information such as the patient's gender, age, medical history, family history, smoking history, pathological staging and typing, and clinical diagnosis. The patient's name, ID number and other information related to the patient's pr...

Embodiment 3

[0181] Embodiment 3, pre-dissociation treatment of gastric cancer tissue samples

[0182] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0183] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0184] 1. Sample weighing.

[0185] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0186] 3. Wash the sample 10 times with sample cleaning solution and 5 times with sterile PBS solution.

[0187] 4. Use ophthalmic scissors, ophthalmic forceps, scalpel and other equipment to carefully peel off the adipose tissue, connective tissue, and necrotic tissue in the sample.

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Abstract

The invention discloses a method for culturing gastric cancer solid tumor primary cells. The invention provides the method for culturing the gastric cancer solid tumor primary cells and an auxiliary reagent. The core of the technology is as follows: (1) treating gastric cancer solid tumor tissues by using a mild cell dissociation reagent, so as to ensure the vitality of cancer cells in the tissuesto the greatest extent; and (2) preparing a special serum-free culture medium, and carrying out in-vitro culture on tumor cells derived from gastric cancer solid tumors by utilizing a suspension culture system, so as to eliminate the interference of normal cells to the greatest extent while the normal amplification of the cancer cells is ensured. The gastric cancer primary cell culture obtained by the method disclosed by the invention can be used for in-vitro experiments of various cell levels, next-generation sequencing, animal model construction, cell line construction and the like. Predictably, the culture method has wide application prospects in the fields of gastric cancer research and clinical diagnosis and treatment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for culturing primary cells of gastric cancer solid tumors. Background technique [0002] Gastric cancer is one of the most common malignant tumors that seriously threaten human health. my country is a country with a high incidence of gastric cancer, and the incidence and death of gastric cancer account for 42.6% and 45% of the global incidence and death respectively. In my country, the incidence rate of gastric cancer is 11.8%, ranking fourth among all malignant tumors. The mortality rate of gastric cancer is 22.0%, ranking fifth among all malignant tumors. With the development of the economy, the improvement of living standards and the change of lifestyle, the incidence of gastric cancer will continue to rise. In addition, gastric cancer has a high risk of recurrence and metastasis, and more than 50% of gastric cancer patients will experience recurrence and metastasis t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/33C12N2501/11C12N2501/115C12N2501/119C12N2500/30C12N2501/30C12N2501/345C12N2501/999C12N2501/998
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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