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Human intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and application thereof

A technique for intrahepatic cholangiocarcinoma and cholangiocarcinoma, which is applied in the field of oncology and cell culture, and can solve the problems of unsatisfactory line establishment, unfavorable in vivo experiments, difficult tumor cell models, etc.

Inactive Publication Date: 2017-09-19
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

[0004] However, it is difficult to obtain an intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and suitable as a tumor cell model. Most of the cells isolated from tumor tissue are not suitable for in vitro culture, the line establishment is not ideal, or the tumorigenic ability is very poor. Poor, difficult to use as a tumor cell model
At present, there are only a few commercial intrahepatic cholangiocarcinoma cell lines included in ATCC in the United States, and even fewer intrahepatic cholangiocarcinoma cell lines established in China, and their tumor formation rates are low, which is not conducive to in vivo experiments

Method used

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  • Human intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and application thereof
  • Human intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and application thereof
  • Human intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1. Obtainment and observation of human intrahepatic cholangiocarcinoma cell line IHC-ST1

[0056] 1. Primary culture

[0057] The IHC-ST1 cells of the present invention come from the liver tumor tissue in situ of a female patient with intrahepatic cholangiocarcinoma. Under sterile conditions, the liver tumor tissue in situ from a female patient undergoing radical resection of intrahepatic cholangiocarcinoma was taken and rinsed in D-Hanks solution (containing 1000 units / ml penicillin, 3 μg / ml amphotericin B) to remove blood stains and necrotic tissue. Then place the tissue in serum-free DMEM / F12 medium and cut it into 1-3mm with ophthalmic scissors 3 Then add 0.5mg / ml type I collagenase and digest at 37°C for 30min. Fetal calf serum was added to stop the digestion, and the tissue block was filtered through a 200-mesh cell sieve along with the soaking solution, and the filtrate was transferred to a centrifuge tube, and centrifuged at 1000 rpm for 5 minutes. T...

Embodiment 2

[0065] Example 2, the growth curve of the human intrahepatic cholangiocarcinoma cell line IHC-ST1 of the present invention

[0066] The human intrahepatic cholangiocarcinoma cell line IHC-ST1 cells in good growth state were taken, digested, made into single cell suspension, and counted. Cells were seeded into 96-well plates with 3000 cells per well. CCK-8 detection was carried out at 12h, 24h, 48h, and 72h after cell inoculation, and the average number and standard deviation of the cell number were calculated, and the cell growth curve was drawn.

[0067] The growth curve obtained is as figure 2 . It can be seen from the figure that the cells grow logarithmically on the 1st to 3rd day, and reach a plateau after the 3rd day, and the doubling time is 24h.

Embodiment 3

[0068] Example 3. Karyotype analysis of the human intrahepatic cholangiocarcinoma cell line IHC-ST1 of the present invention

[0069] Cells in exponential growth phase, 80%-90% confluent monolayer culture were taken. Add colchicine to suppress metaphase division, so that the final concentration in the medium is 0.04-0.1μg / ml medium, CO 2 Continue culturing for 4 hours in the incubator.

[0070] After fixed staining, the chromosomes of 30 cells in metaphase were counted. The number of chromosomes in the IHC-ST1 cell line was mainly concentrated in 55-70, and the chromosome mode was 62, suggesting the existence of hyperdiploidy. See the representative picture image 3 .

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Abstract

The invention relates to a human intrahepatic cholangiocarcinoma cell line with high tumorigenic ability and application thereof. The invention discloses a novel human intrahepatic cholangiocarcinoma cell line which grows stably and has clear quantity of generations and high tumorigenic rate; the human intrahepatic cholangiocarcinoma cell line can form an adenoid tumor in vivo and can be used for in-vivo and in-vitro research on human intrahepatic cholangiocarcinoma; a foundation is laid for constructing an in-vivo animal model, disclosing a tumor malignant progression mechanism, screening related biomarkers and developing novel intrahepatic cholangiocarcinoma resisting medicines.

Description

technical field [0001] The invention belongs to the field of oncology and cell culture, and more specifically, the invention relates to a human intrahepatic cholangiocarcinoma cell line with high tumor forming ability and application thereof. Background technique [0002] Intrahepatic cholangiocarcinoma (ICC) refers to a malignant tumor originating from bile duct epithelial cells above the confluence of the left and right hepatic ducts. It has high malignancy, hidden symptoms and poor prognosis. In recent years, the morbidity and mortality of ICC have gradually increased, and its main risk factors include intrahepatic bile duct stones, viral hepatitis, cholangitis, and parasitic infections. Surgical resection is still the most important method for the treatment of ICC, but due to the difficulty of early diagnosis, only 10% of patients can undergo radical resection, and most patients have reached an advanced stage when they are diagnosed and cannot be operated. The adjuvant ...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
CPCC12N5/0693G01N33/5011
Inventor 王红阳付静谈冶雄胡丛莉孙焕林
Owner HEPATOBILIARY SURGERY HOSPITAL SECOND MILITARY MEDICAL UNIV
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