Liver cancer cell system from liver cancer portal vein tumor thrombus of human and system establishing method thereof

A technology of liver cancer cell line and portal vein, applied in the field of liver cancer cell line and its establishment

Inactive Publication Date: 2009-12-09
HEPATOBILIARY SURGERY HOSPITAL SECOND MILITARY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the liver cancer cell line and its estab...

Method used

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  • Liver cancer cell system from liver cancer portal vein tumor thrombus of human and system establishing method thereof
  • Liver cancer cell system from liver cancer portal vein tumor thrombus of human and system establishing method thereof
  • Liver cancer cell system from liver cancer portal vein tumor thrombus of human and system establishing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Establishment of HCC cell line derived from portal vein tumor thrombus of human HCC

[0027] 1. Materials

[0028] 1. Two different cell culture media and their preparation methods

[0029] 1.1 Culture solution D / F: DMEM / F12 (1:1) (GIBCO, USA), 10% (v / v) fetal bovine serum (Biochrom AG, USA), EGF (Epidermal Growth Factor, epidermal growth factor) 20ng / ml (U.S. Sigma Company), 5ug / ml insulin, 0.30g / L L-glutamine, 100U / L penicillin, 100ug / L streptomycin;

[0030] 1.2 Culture solution DMEM: high-sugar DMEM medium (GIBCO, USA), 10% (v / v) fetal bovine serum (Biochrom AG, USA), 0.30g / L L-glutamine, 100U / L penicillin, 100ug / L Streptomycin.

[0031] 2. Other cell culture materials

[0032] Cell sieve (BD Company, U.S.); program freezing box (NALGENE Company, U.S.); 0.05% type I collagenase solution (Sigma Chemical Company, St.Louis, MO); trypsin digestion solution: 0.25% trypsin (GIBCO, U.S. company)+0.02% EDTA, pH value is 7.3; cell cryopreservation solution (DMEM:FBS:D...

Embodiment 2

[0054] Biological characteristics of liver cancer cell lines derived from portal vein tumor thrombus of human liver cancer

[0055] In this example, conventional methods were used to detect the biological characteristics of the cell line CSQT-1 with the deposit number CCTCC NO: C200814. Methods as below:

[0056] 1. Experimental method

[0057] 1. Morphology

[0058] (1) General shape

[0059] Two days after cell inoculation, the growth morphology of the population cells was observed.

[0060] (2) light mirror

[0061] The morphological structure and growth characteristics of the cultured living cells were observed under an inverted phase-contrast microscope.

[0062] (3) electron microscope

[0063] The 40th passage cells were taken, and the ultrastructure of the cells was observed by scanning and transmission electron microscopy.

[0064] 2. Cell Kinetics

[0065] (1) Drawing of cell growth curve and cell population doubling time

[0066] Take well-grown logarithmic...

Embodiment 3

[0111] Culture medium D / F (1)

[0112] DMEM / F12 (1:1) (GIBCO, USA)

[0113] 10% (v / v) fetal bovine serum (Biochrom AG, USA)

[0114] EGF (Sigma, USA) 20ng / ml

[0115] Insulin 5ug / ml

[0116] L-Glutamine 0.30g / L

[0117] Penicillin 100U / L

[0118] Streptomycin 100ug / L

[0119] Preparation method: The culture medium D / F can be prepared according to the above concentration.

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Abstract

The invention relates to the field of microorganism animal cells, in particular to a liver cancer cell system from liver cancer portal vein tumor thrombus of human and a system establishing method thereof. The invention adopts the following technical scheme that the liver cancer cell system from liver cancer portal vein tumor thrombus of human is preserved in the China Center for Type Culture Collection (CCTCC) and has a preservation number of CCTCCNO: C200814. The system establishing method of the liver cancer cell system from liver cancer portal vein tumor thrombus of human comprises the following steps: (1) acquiring cancer embolus cells and carrying out a primary culture on the embolus cells; (2) carrying out a clone expansion culture on a tumor cell island; and (3) carrying out a continuous culture on tumor cells. The invention has the advantages that CSQT-1 cells are obtained from the portal vein tumor embolus and have incomparable advantages on the forming mechanism and the tendency growing mechanism of the portal vein tumor embolus than other existing liver cancer systems. The cell system has wide prospect of application, is a sub-hypotetraploid displayed by the karyotype analysis and accounts for 58 percent of the total.

Description

technical field [0001] The invention relates to the field of microbial animal cells, in particular to a liver cancer cell line derived from portal vein tumor thrombus of human liver cancer and a method for establishing the line. Background technique [0002] Primary liver cancer (HCC) is one of the most common malignant tumors, and the median survival time of untreated patients is less than one year. Surgical resection is currently the most effective treatment for liver cancer. But even if the tumor is removed, most patients still die of postoperative tumor recurrence and metastasis. The main feature of liver cancer metastasis is to grow into the portal vein to form portal vein tumor thrombus. Shanghai Oriental Hepatobiliary Surgery Hospital performed liver tumor resection on 5,524 patients with liver cancer, and the proportion of portal vein tumor thrombi found in the resected specimens reached 67.1% (Chinese Journal of Surgery, 2001, 39: 25-28); The study of growth char...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12R1/91
Inventor 程树群胡华生吴孟超
Owner HEPATOBILIARY SURGERY HOSPITAL SECOND MILITARY MEDICAL UNIV
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