Human hepatocellular carcinoma cell line, and establishing method and application thereof

A method for establishing a method and a technology for cancer cells, applied in the field of cell lines, can solve the problems of large differences in biological characteristics of liver cancer and low biodiversity, and achieve the effect of weight loss and stable characteristics

Pending Publication Date: 2015-07-01
ZHONGSHAN HOSPITAL FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a new human hepatocellular carcinoma cell line in view of the existing human hepatocellular carcin

Method used

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  • Human hepatocellular carcinoma cell line, and establishing method and application thereof
  • Human hepatocellular carcinoma cell line, and establishing method and application thereof
  • Human hepatocellular carcinoma cell line, and establishing method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Preparation of LIXC-012 cells

[0043] SCID mice: 5 SCID mice, male, weighing 16.0±1.0 g, aged 5 weeks, were raised in SPF environment. SCID mice were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

[0044] A fresh resection specimen of clinical liver cancer was obtained from Shanghai Zhongshan Hospital (male, 71 years old, primary liver cancer, pathological diagnosis: (right lobe of liver) hepatocellular carcinoma, grade II differentiation, nodular cirrhosis of surrounding liver tissue, pulse Tumor embolus in the tube), immediately immersed in pre-cooled sterile HBSS buffer (containing 500 U / mL penicillin G, 500 μg / mL streptomycin sulfate, and 1.25 μg / mL amphotericin B). In the biosafety cabinet, the specimen was washed with fresh sterile HBSS buffer solution, cut into small pieces of 20-50 mg, and punctured and inoculated subcutaneously in the axillae of SCID mice. 30 days after inoculation, small tumor nodules were found under the skin o...

Embodiment 2

[0049] Biological characteristics and application of LIXC-012 cells

[0050] The invention adopts RPMI1640 culture solution containing fetal bovine serum and insulin to cultivate LIXC-012 cells, so that they can grow and pass stably in vitro for a long time. When the cells are passed to more than 20 generations, the cell properties are gradually stabilized, and the relevant biology, genetics and tissue source identification are carried out until the 50th generation has the same stable properties. Through experimental observation and verification, the LIXC-012 cells grown in vitro are epithelial-like. Under an inverted microscope, the cells adhere to the wall, disperse uniformly, vary in size, and have a cobblestone-like shape. The cells have a typical epithelial-like morphology and lose contact growth inhibition. , showing malignant growth. Genetic studies have confirmed that the cells are aneuploid, with severe chromosome number and structural aberrations, consistent with th...

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Abstract

The invention discloses a human hepatocellular carcinoma cell line, and an establishing method and application thereof. The human hepatocellular carcinoma cell line is preserved in China Center for Type Culture Collection with an accession number of CCTCC No: C201114. The establishing method comprises the following steps: 1) acquiring a fresh clinical liver cancer excision specimen, cutting the specimen into small blocks with sizes in a range of 20 to 50 mg and inoculating mammals with the small blocks; and 2) in 70 to 90 d after inoculation, killing tumor-bearing mammals, taking out tumor tissue and carrying out primary culture and subculturing of cancer cells. The human hepatocellular carcinoma cell line provided by the invention has stable properties and tumorigenicity, can realize stable several passages and successful preparation of a liver cancer animal model, can be used for analyzing in-vivo and in-vitro drug susceptibility so as to further realize establishment of mutually-related in-vivo and in-vitro drug screening platforms, and is an ideal human primary hepatic carcinoma cell line applied to fundamental research and preclinical application.

Description

technical field [0001] The invention relates to the field of cell lines, in particular to a human hepatocellular carcinoma cell line and its establishment method and application. Background technique [0002] Primary liver cancer is a common malignant tumor with the characteristics of high incidence, poor prognosis and high mortality. According to the latest statistics, there are about 748,000 new cases of liver cancer worldwide, and 696,000 deaths, half of which occurred in China. Its incidence and death rate rank fifth and second among male tumor patients, and rank seventh and sixth among female tumor patients. [0003] The onset of primary liver cancer is hidden, lack of typical symptoms in the early stage, early diagnosis is difficult, and the course of the disease develops rapidly. Once the symptoms are obvious, most of the patients who seek medical treatment on their own are in the middle and late stages. Patients with advanced liver cancer often have symptoms such a...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027A61K49/00C12Q1/02C12R1/91
Inventor 代智许春花樊嘉周俭胥甜甜胡刚吴伟忠汤旭蓁秦宵然
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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