Method for constructing pancreatic cancer cell line by positive expression of Chinese B7-H5

A technology of sipanc-1076 and B7-H5, which is applied in the field of pancreatic cancer primary tumor cell line establishment, can solve the problems of application limitations and achieve good tumorigenicity

Inactive Publication Date: 2019-02-12
梁廷波
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Problems solved by technology

Due to the lack of blood supply of pancreatic cancer itself [5] , and many tumor-associated fibroblasts (cancer associated fibroblasts, CAF) e

Method used

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  • Method for constructing pancreatic cancer cell line by positive expression of Chinese B7-H5
  • Method for constructing pancreatic cancer cell line by positive expression of Chinese B7-H5
  • Method for constructing pancreatic cancer cell line by positive expression of Chinese B7-H5

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Embodiment Construction

[0015] The present invention will be described in detail below in conjunction with accompanying drawing:

[0016] The method for building a system of the present invention:

[0017] 1. Specimen source: The specimen was taken from a 67-year-old male patient in the Department of Hepatobiliary and Pancreatic Surgery of the Second Affiliated Hospital of Zhejiang University School of Medicine. Pathological diagnosis: pancreatic ductal adenocarcinoma, TNM stage: T 3 N 0 m 0 .

[0018] 2. Organoid organoid complete medium configuration: AdDMEM / F12 medium, 10mMHEPES, 2mMGlutamax, penicillin / streptomycin double antibody 100mg / ml, 1 times B27 (b27 is a mixture of many formulas, the concentration of the manufacturer is the working concentration 100 times, diluted to 1 times when used), 1mg / ml primary cell antibiotic (primocin), 1mM N-acetylcysteine, 50% (v / v) Wnt3a conditioned medium, 10% (v / v ) RSPO1 conditioned medium, 10% (v / v) Noggin conditioned medium, 50ng / ml epidermal growth f...

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Abstract

The invention discloses a method for constructing a pancreatic cancer cell line by positive expression of Chinese B7-H5. The method comprises the following steps of 1, collecting of a specimen; 2, preparing of an Organoid type organ complete culture medium; 3, digestion of tissues; 4, constructing and culture of Organoid type organs of pancreatic cancer; 5, cell generation pass and culture. The method has the beneficial effects that the biology features of the constructed Chinese pancreatic cancer cell line sipanc-1076 are used as pancreatic ductal adenocarcinoma; the particular characteristicof positive expression of B7-H15 which does not exist in other commercialized human pancreatic cancer cell lines is realized; the tumor forming property is good; the method can be applied to the study on tumor immune escape of pancreatic cancer, analysis of related genealogy of Chinese pancreatic cancer, and the like.

Description

technical field [0001] The invention relates to a method for establishing a primary tumor cell line of pancreatic cancer, mainly a method for establishing a Chinese B7-H5 expression-positive pancreatic cancer cell line sipanc-1076. Background technique [0002] Pancreatic ductal adenocarcinoma (Pancreatic ductal adenocarcinoma, PDAC) is the most common pancreatic tumor. Metastasis occurs in the early stage, but the symptoms are not obvious. When it is discovered, most of them are in the advanced stage, and the prognosis is extremely poor. The five-year survival rate after diagnosis is often less than 5%. Known as the "King of Cancer" [1-3] , ranking sixth in the death rate of male cancer patients in my country [4] . Most of the pancreatic cancer cell lines used in basic and clinical research in the past came from foreign ATCC cell banks, such as Panc-1, Bxpc-3, Miapaca-2, ASPC-1, etc., and the cell lines were derived from Caucasian pancreatic cancer patients , for pancreat...

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Application Information

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IPC IPC(8): C12N5/09C12R1/91
CPCC12N5/0693
Inventor 梁廷波白雪莉王健鑫陈琦魏涛王艺倪磊王建锋
Owner 梁廷波
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