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Chinese lung adenocarcinoma cell system and application thereof

A technique for lung adenocarcinoma cells and lung adenocarcinomas, applied in the field of microbial medicine, can solve problems such as ineffectiveness, and achieve the effect of promoting proliferation

Active Publication Date: 2017-08-22
SHANGHAI CHEST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most lung cancers do not have gene mutations such as EGFR, and targeted drugs have little effect on them. Traditional chemotherapy and radiotherapy are still the main treatment for this type of lung cancer

Method used

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  • Chinese lung adenocarcinoma cell system and application thereof
  • Chinese lung adenocarcinoma cell system and application thereof
  • Chinese lung adenocarcinoma cell system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Isolation and processing of primary tumor cells

[0023] Fresh Chinese tumor tissue blocks obtained from the operating table of the hospital were isolated and cultured for primary tumor cells in a sterile environment.

[0024] 1. First put the tumor tissue block into a 50ml centrifuge tube, shake and wash 5 times with PBS containing 10 times the double antibody until there is no blood. Place the tissue block in a Petri dish and remove excess tissue, such as fat or dead tissue.

[0025] 2. Move the tissue to another plate, and use a cross scalpel to cut it into about 1mm 3 The size of the organization block. Take a 70um filter and place it on a 50ml centrifuge tube. Gently pressurize the core of the syringe to force the tissue through the mesh into the culture medium, draw the culture medium and blow it through the mesh to flush the cells down.

[0026] 3. Add 5ml of trypsin / collagenase II to the culture dish of the remaining tissue pieces, digest in a 37°C incubator...

Embodiment 2

[0032] Example 2. Monoclonal culture of primary tumor cells

[0033] 1. Digest the cells with trypsin to prepare a single cell suspension, count the cells, and dilute the cells to 1×10 cells / mL. The specific steps are as follows:

[0034] a) Dilute the digested cells to 1×10 5 individual / mL.

[0035] b) Take 1×10 5 cells / mL cell suspension 200uL, add to 20mL, become 1×10 3 individual / mL.

[0036] c) Divide 1×10 3 Cells / mL cell suspension 200uL, add to 20mL, become 1×10 cells / mL.

[0037] 2. Add 100uL of 1×10 cells / mL cell suspension to the microtiter culture plate of 96-well plate, and one colony will be formed in each well.

[0038] 3. A few hours after inoculation, find the culture well with only one cell through the microscope. The wells were marked and followed, and a drop in pH indicated cell growth, confirmed by microscopic observation. After the cells were overgrown, trypsinization was inserted into a 6-well plate for subculture.

Embodiment 3

[0039] Example 3. Detection and identification of Chinese lung adenocarcinoma cell line CAFQ1

[0040] 1. Cytomorphological observation of primary tumor cell lines

[0041] Under the light microscope, most of the tumor cells were spindle-shaped and oval-shaped, and a few were polygonal. The two poles of the cells were pointed, and the nucleus was obvious. The cells in the logarithmic growth phase spread well, and CAFQ1 randomly spread, and most of them were oval epithelioid cells, polygonal, and more regular in shape. Such as figure 1 shown.

[0042] 2. Immunofluorescence detection of cell marker proteins

[0043] 1) After trypsinizing the cells, suspend them in 1mL of complete culture medium, pipette fully to form a single-cell suspension, and perform cell counting. Insert 5×10 into 24-well plate as needed 4 cells. Before adding the cell suspension, add a small amount of medium to each well to make the slide and the culture dish stick together to reduce the entry of cel...

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Abstract

The invention discloses a Chinese lung adenocarcinoma cell system CAFQ1 and application thereof. The preservation number of the cell system is CCTCC NO: C201774. The cell system has similar protein expression as that of primary tumor tissue, also has chromosome abnormal karyotype and has tumor properties of in-vitro tumor formation properties. By adopting the cell system, properties of lung adenocarcinoma tumor cells can be maintained after continuous in-vitro subculture, and the cell system is applicable to cell materials for researching and developing immunity treatment, diagnosis and medicines for lung adenocarcinoma. The cell system disclosed by the invention comprises ALK: Kras mutations, the two mutations can result normal pulmonary epithelial cell mutations and tumor formation and can promote tumor cell proliferation, therefore, the cell system can be used as powerful tool cells for studying target medicines with ALK; Kras as targets.

Description

technical field [0001] The present invention relates to the field of microbiology medicine, and more specifically relates to a Chinese lung adenocarcinoma cell line CAFQ1 and its application. Background technique [0002] Lung cancer is one of the tumors with the highest mortality rate in the world, accounting for 19.4% of the deaths caused by tumors. In Europe, with the reduction of the number of smoking in China and the increase of female smokers, the incidence rate of men is decreasing year by year, and the incidence rate of women is increasing year by year. Among them, 80%-85% are non-small cell lung cancer, and the 5-year survival period is low at 11%. The epidemiological study of lung cancer in Asia in 2014 showed that 43.4% of lung cancers were females, the average age of onset was 60 years old, 52.6% had no smoking history, and 51.4% had EGFR mutations. Lung cancer is mainly divided into non-small cell lung cancer and small cell lung cancer. Non-small cell lung ca...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12Q1/68G01N33/574C12R1/91
CPCC12N5/0693C12N2509/00C12Q1/6886C12Q2600/156G01N33/5011G01N33/5026G01N33/57423G01N33/57484
Inventor 路丽明孙攀郑宁张杰赵丹丹杨玉琴
Owner SHANGHAI CHEST HOSPITAL
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