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Human biliary tract cancer cell lines and application

A cancer cell and biliary tract technology, which is applied to human biliary tract cancer cell lines and application fields, can solve the problems of lack of in vitro culture system and poor therapeutic effect of biliary tract cancer.

Inactive Publication Date: 2018-05-18
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, one of the main reasons for the poor treatment effect of biliary tract cancer is the lack of a perfect in vitro culture system.

Method used

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  • Human biliary tract cancer cell lines and application
  • Human biliary tract cancer cell lines and application
  • Human biliary tract cancer cell lines and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Samples were taken from in situ tumor tissues of a 69-year-old male patient with gastric cancer and gallbladder cancer, a 66-year-old sexual patient with hilar cholangiocarcinoma, and a 57-year-old female patient with hilar cholangiocarcinoma. Wash with normal saline 3 times. Put the tissue into a sterile petri dish, drop a little collagenase digestion solution on the surface of the tissue block, and use different sterile small ophthalmic scissors to fully cut it to a volume of about 1-2mm 3 Size: Collect the chopped tumor tissue with collagenase digestion solution and put it in a 15ml centrifuge tube, add a certain amount of digestion solution, and put it in 37°C CO 2 Digest overnight in an incubator; centrifuge the digested tissue and cell suspension overnight (300×g, 7min), discard the supernatant, wash and centrifuge 3 times with normal saline; resuspend sediment in RPMI 1640+10% FBS+ Place in a Petri dish at 37°C CO 2 Culture in an incubator, do not move the cult...

Embodiment 2

[0045] Sample preparation: 1 x 10 7 The cultured ZJU-0430, ZJU-0826 and ZJU-1125 cells were trypsinized and collected in a 15ml centrifuge tube, and the cells were collected by centrifugation and then washed and centrifuged again;

[0046] Fixation: add 2.5% glutaraldehyde fixative to the cells for 2 hours, wash with 0.1M phosphate buffer for 3 times, add 1% glutaric acid fixative for 2 hours, and wash with 0.1M phosphate buffer for 3 times;

[0047] Dehydration: 50% ethanol → → 70% ethanol → → 90% ethanol → → 90% ethanol + 90% acetone (1:1) → → 90% acetone → → 100% acetone (3 times) in a refrigerator at 4°C , each process is maintained for 15-20 minutes;

[0048] Embedding: pure acetone + embedding solution (2:1) at room temperature for 3-4 hours → → pure acetone + embedding solution (1:2) overnight at room temperature → → pure embedding solution at 37°C for 2-3 hours;

[0049] Curing: Overnight in 37°C oven → → 12h in 45°C oven → 24h in 60°C oven;

[0050] Preparation of ...

Embodiment 3

[0055] After the cultured ZJU-0430, ZJU-0826 and ZJU-1125 cells were trypsinized, resuspended in RPMI 1640+10% FBS medium and counted, the final concentration was adjusted to 2.5×10 4 / ml and 5×10 4 / ml of cell suspension; inoculate them in 96-well plates, 200 μl per well, set 6 duplicate wells in each group, and the wells with only medium as blank control; put the culture plate into a 37 ° C incubator for culture , CCK-8 was added at 0, 12, 24, 36, 48, 60 and 72h and incubated for 2.5h, then the absorbance was measured at 450nm. The result is as image 3 As shown, the population doubling time is about: 24h, 24h and 48h, image 3 Middle: A: ZJU0430; B: ZJU-0826; C: ZJU-1125.

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Abstract

The invention belongs to the field of microbe animal cell lines, and particularly relates to human biliary tract cancer cell lines and application. The human biliary tract cancer cell lines comprise human biliary tract cancer cell lines which are called a human gallbladder carcinoma cell line ZIJ-0430, a human hilar cholangiocarcinoma cell line ZJU-0826 and a human hilar cholangiocarcinoma cell line ZJU-1125 respectively, and the three human biliary tract cancer cell lines are conserved in the China Typical Culture Conservation Center with the conservation number of CCTCC NO: C2017174, CCTCC NO:C2017175 and CCTCC NO:C2017176 respectively. The human biliary tract cancer cell lines ZJU-0430, ZJU-0826 and ZJU-1125 are built from the source of Chinese people, the line building time is short, the biological inheritance trait is stable, and as study models, the biliary tract cancer cell lines provide great help for knowing a primary biliary tract cancer pathogenesis of Chinese people.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to human biliary tract cancer cell lines and applications. Background technique [0002] Despite advances in diagnostic and therapeutic techniques, the survival prognosis of patients with biliary tract cancer is poor. Previous studies have reported that surgical resection is the only way to treat the disease, and there is no effective chemotherapy regimen for patients without surgery or postoperative recurrence. Exploring the nature of this tumor biology is extremely important to improve the prognosis of patients with biliary tract cancer. At the same time, one of the main reasons for the current poor treatment effect of biliary tract cancer is the lack of a perfect in vitro culture system. However, currently in vitro cultured tumor cell lines play an irreplaceable role in the research of tumorigenesis, development, invasion, metastasis mechanism, early diagn...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027C12R1/91
CPCA01K67/027A01K2207/12A01K2227/105A01K2267/0331C12N5/0693
Inventor 孙继红杨晓明周飞张艳华
Owner ZHEJIANG UNIV
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