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Establishment and application of hepatocellular carcinoma cell line HCC-LY10

A HCC-LY10, liver cancer cell technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of poor tumorigenicity, non-expression of markers, unsatisfactory and other problems

Inactive Publication Date: 2012-09-26
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For the field of liver cancer, at present, although there are some liver cancer cell lines, these liver cancer cell lines have many disadvantages, including: unstable passage, poor tumorigenicity, inconsistent characteristics of cell lines, expression of liver cancer-specific markers, etc. Irregular, some markers are not even expressed, etc., so it is not satisfactory

Method used

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  • Establishment and application of hepatocellular carcinoma cell line HCC-LY10
  • Establishment and application of hepatocellular carcinoma cell line HCC-LY10
  • Establishment and application of hepatocellular carcinoma cell line HCC-LY10

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Establishment of hepatocarcinoma cell line HCC-LY10

[0068] (a) source

[0069] The cell line comes from a male liver cancer patient (patient number: 604648), and its clinical information is shown in the table below.

[0070] Table 1

[0071]

[0072] (b) method

[0073] The primary liver cancer tissue of the liver cancer patient was inoculated into immunodeficient mice, and the tumor tissue was taken for repeated inoculation and passage, and finally a stable passage cell line was obtained. Specifically include the following processes:

[0074]

[0075]

[0076] The tumor-bearing NOD / SCID mice were sacrificed painlessly, and the subcutaneous tumors were aseptically extracted. Place the tumor in a pre-cooled PBS culture dish, fully clean up the tumor capsule and necrotic part of the tumor, use ophthalmic scissors to cut the tumor tissue into small pieces with a size of about 5×5×5 mm, and add a small amount of liver cells The culture medium is cultivated ...

Embodiment 2

[0081] Biological characteristics of human hepatoma cell line HCC-LY10

[0082] In this example, the biological characteristics of the purebred human liver cancer cell line HCC-LY10 with the deposit number CCTCC NO: C2010100 were detected by conventional methods. The result is as follows:

[0083] 2.1 Cell morphology:

[0084] Observation of HCC-LY10 liver cancer cells under an inverted phase-contrast microscope reveals that the cells are polygonal or round, uniform in shape, multinucleated giant cells can be seen, and the cell proliferation speed is fast, which can be passaged every 48 hours ( image 3 ).

[0085] 2.2 Enzyme / GFP double labeling

[0086] The results of luciferase / GFP double labeling of HCC-LY10 cell line are as follows Figure 4 (right panel), indicating that the cells were all labeled with green fluorescent protein (GFP).

[0087] 2.3 Karyotype analysis

[0088] By adding colchicine (final concentration: 0.04 μg / ml) to the cell culture medium, a large n...

Embodiment 3

[0115] Establishment of Animal Model of Liver Cancer Using Human Hepatoma Cell Line

[0116] Take the human liver cancer cell line HCC-LY10 (preservation number: CCTCC C2010100) obtained in Example 1, digest it with the digestive solution (0.25% trypsin, 0.02% EDTA, pH7.3), collect and count, and count as 1×10 6 or 1×10 7 Number of cells / mouse Inoculate subcutaneously on the right chest wall of 20 Balb / c nude mice aged 4-6 weeks.

[0117] Between 14 and 60 days after inoculation, tumors appeared in all 20 mice. Pathological detection was carried out by the same method as in Example 2, and it was confirmed that hepatocellular carcinoma was induced in the animal model.

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Abstract

The invention provides establishment and application of a hepatocellular carcinoma cell line HCC-LY10. Specifically, the hepatocellular carcinoma cell line HCC-LY10 provided in the invention has stable traits, can be subcultured stably and repeatedly, and is applicable to established animal models of hepatocellular carcinoma, thus being an ideal cell line for the basic and preclinical phase application study of hepatocellular carcinoma. In addition, the hepatocellular carcinoma cell line HCC-LY10 partially expresses hepatocellular carcinoma stem cell marker EpCAM. The invention also provides application of the cell line in establishing model animals, screening drugs, and other aspects.

Description

technical field [0001] The present invention relates to the fields of biology and oncology, and more specifically relates to a new liver cancer cell line HCC-LY10, its establishment method and application. Background technique [0002] The occurrence and development of various human diseases are very complicated, and in-depth exploration of the pathogenesis and therapeutic mechanism of various diseases cannot be carried out on patients. However, it is possible to follow the Animal Protection Law, conduct research on various diseases and life phenomena through animals, and then deduce it to human beings to explore the mysteries of human life and the laws of the occurrence and development of human diseases, so as to control the occurrence and development of human diseases, cure diseases and Reduce the suffering of human beings, improve the quality of life, and prolong the life of human beings. [0003] For a long time, people have found that it is difficult to promote the dev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A61D7/00A01K67/027A61K49/00
Inventor 李锦军姚明闫明霞张立行刘蕾葛超郝向芳顾健人
Owner SHANGHAI INST OF ONCOLOGY
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