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Human liver cancer cell line of high lymphatic channel tranfer

A technology of human liver cancer cell line and lymph node metastasis, which is applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., and can solve the problem of lack of experimental platform for liver cancer lymphatic metastasis

Inactive Publication Date: 2004-11-17
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, looking at the human liver cancer models at home and abroad, there is no human liver cancer cell line with clear lymphatic metastasis potential, and the research on lymphatic metastasis of liver cancer lacks a good experimental platform

Method used

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  • Human liver cancer cell line of high lymphatic channel tranfer
  • Human liver cancer cell line of high lymphatic channel tranfer

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Example 1 Establishment of a human liver cancer cell line with high lymphatic metastasis potential

[0018] 1. Inoculate the subcutaneous part of the paw pad of nude mice: take 3×10 HCCLM6 cells 6 / 0.05ml inoculated in the subcutaneous part inside the right paw pad of nude mice. Nude mice are BALB / C-nu / nu, male, 4-6 weeks old, purchased from Shanghai Institute of Materia Medica, Chinese Academy of Sciences, and raised in a special pathogen-free environment. After 5 weeks, aseptically remove the metastatic lymph nodes from the popliteal fossa, groin, common iliac, and renal hilus on the same side of the nude mice.

[0019] 2. In vitro primary culture and expansion of tumor cells in the metastatic lymph nodes: put the above-mentioned metastatic lymph nodes in a sterile plate, wash with PBS, remove the connective tissue outside the lymph nodes, grind on a 200-mesh stainless steel screen, and use 10% (v / v) Fetal bovine serum (Hyclone, Utah, USA) was washed with high-suga...

Embodiment 2

[0024] Nude mouse model of liver orthotopic transplantation of HCCLYM cell line

[0025] 1. The conventional culture conditions in vitro are high-glucose DMEM medium (GIBCOBRL, Grand Island, NY, USA) containing 10% (v / v) fetal bovine serum (Hyclone, Utah, USA), 37°C, 5% CO 2 / 95% air, saturated humidity.

[0026] 2. HCCLM3 cells grown to the logarithmic growth phase were routinely digested with 0.25% trypsin + 0.1% EDTA, the cells were washed with PBS, the viable cells were counted by trypan blue exclusion method, and the cell suspension was prepared at a concentration of 2.5×10 7 / ml.

[0027] 3. Take 0.2 ml of the above cell suspension (5×10 6 1), inoculated subcutaneously in the back near the axilla of 4-6 week-old BALB / C-nu / nu male nude mice, and reared in an environment free of special pathogens.

[0028] 4. After 2 weeks, the subcutaneous tumor grew to a diameter of about 1.0 cm, and the subcutaneous tumor was cut into small pieces of tumor tissue with a diameter of 1...

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Abstract

The invention relates to the field of microbiological animal cells, in particular to a human hepatocarcinoma cell line with high lymph channel transfer potency established in lymph gland transfer stove, wherein nude mouse in vivo continuous screening method lymph channel transfer screening is employed to create human liver cancer cell line HCCLYM with high lymphatic channel transfer proficiency.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to establishing a human liver cancer cell line with high lymphatic metastasis potential from lymph node metastases. Background technique [0002] Since the 1990s, primary liver cancer has become a malignant tumor with the second highest mortality rate in my country. Primary liver cancer is highly malignant and progresses rapidly. Surgical resection is the most effective treatment. However, even for small liver cancers with a diameter less than 5 cm, the 5-year recurrence rate after surgery is as high as 40-50%. Postoperative recurrence and metastasis is the biggest obstacle to the survival of liver cancer patients. The main ways of liver cancer metastasis are hematologic metastasis, lymphatic metastasis, direct invasion and implantation. Clinical practice has shown that lymphatic metastasis is not uncommon in patients with liver cancer. Kojiro et al. conduct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 田波汤钊猷
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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