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Culture medium for culturing primary cells related to cholecystic cholangiocarcinoma

A technology of primary cells and culture medium, applied in the biological field, can solve the problems of long culture period, difficult to remove miscellaneous cells, low culture success rate, etc., and achieve the effect of short culture period, high cell purity and high culture stability.

Inactive Publication Date: 2021-05-07
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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  • Culture medium for culturing primary cells related to cholecystic cholangiocarcinoma
  • Culture medium for culturing primary cells related to cholecystic cholangiocarcinoma
  • Culture medium for culturing primary cells related to cholecystic cholangiocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1. Preparation of reagents for culturing gallbladder cancer, cholangiocarcinoma solid tumor primary cells

[0069] 1. Sample preservation solution (100mL)

[0070] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0071] Table 1 Sample Preservation Solution (100mL)

[0072]

[0073] 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.

[0074] 2. Sample cleaning solution (100mL)

[0075] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0076] Table 2 Sample cleaning solution (100mL)

[0077]

[0078] The sample cleaning solution should be prepared and used immediately.

[0079] 3. Sample dissociation solution (10mL)

[0080] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0081] Table 3 Sample Dissociation Solution (10mL) ...

Embodiment 2

[0173] Example 2. Acquisition of gallbladder cancer and cholangiocarcinoma postoperative specimens / biopsy puncture specimens

[0174] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0175] 2. The attending physician 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 for surgical samples were: primary gallbladder cancer or cholangiocarcinoma, pathological stage II, III or IV, metastatic lesions of gallbladder cancer or cholangiocarcinoma of various pathological types, and surgical specimens weighing more than 20 mg. The selection criteria for biopsy puncture samples are: primary gallbladder cancer or cholangiocarcinoma, pathological stage II, III or IV, metastatic lesions of gallbladder cancer or cholangiocarcinoma of various pathological type...

Embodiment 3

[0179] Example 3, pre-dissociation treatment of gallbladder cancer and cholangiocarcinoma samples

[0180] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0181] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0182] 1. Sample weighing.

[0183] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0184] 3. Wash the sample 10 times with sample cleaning solution and 5 times with sterile PBS solution.

[0185] 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 culture medium for culturing primary cells related to cholecystic cholangiocarcinoma. The culture medium for culturing primary cells related to cholecystic cholangiocarcinoma is a special serum-free culture medium. The culture medium is used for carrying out in-vitro suspension culture on tumor cells derived from cholecystic cholangiocarcinoma, so that the interference from normal cells can be eliminated to the greatest extent while normal amplification of cancer cells is ensured. The primary cell culture of cholecystic cholangiocarcinoma obtained by the method can be used for various cellular in-vitro experiments, next-generation sequencing, construction of animal models, construction of cell lines and the like. Predictably, the culture method has a wide application prospect in the fields of research and clinical diagnosis and treatment of the cholecystic cholangiocarcinoma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture medium for culturing primary cells of gallbladder and cholangiocarcinoma. Background technique [0002] Gallbladder and cholangiocarcinoma is a common malignant tumor of the digestive system that occurs in the gallbladder, bile duct, and intrahepatic bile duct, including gallbladder cancer and cholangiocarcinoma. The total incidence of gallbladder and bile duct-related malignant tumors in my country is about 3%, of which cholangiocarcinoma accounts for 2%, ranking fifth among malignant tumors of the digestive tract in my country. Although its incidence rate is not high, gallbladder and bile duct-related cancers are extremely malignant. Cholangiocarcinoma is even called "the king of liver cancer" and "the king of cancers". For unresectable gallbladder cancer, the median survival is only 8 months. [0003] Although scientific research and medical institutions all over the w...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/02A01N1/02
CPCC12N5/0693A01N1/0221C12N2501/11C12N2501/115C12N2501/12C12N2501/119C12N2501/415C12N2501/01C12N2500/38C12N2500/25C12N2500/46C12N2501/392C12N2501/998C12N2501/71C12N2500/34C12N2501/999C12N2500/36C12N2501/395C12N2501/405
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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