Cis-platinum drug-resistant lung cancer organoid, culture method thereof, medium used for culture of cis-platinum drug-resistant lung cancer organoid and application of cis-platinum drug-resistant lung cancer organoid

A culture medium and organoid technology, applied in the field of biomedicine, can solve problems such as time-consuming, difficult to control cell state, and widely different data

Active Publication Date: 2020-12-15
中关村科技租赁股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Drug concentration escalation and continuous action method usually takes 6 months or more to induce drug resistance, which takes a long time, and the mode of action of its chemotherapy drugs is significantly different from the principle of high-dose and short-course chemotherapy in clinical practice
The high-dose drug intermittent induction method induces drug resistance when the drug concentration is high, and the cells are often difficult to tolerate the sudden change of the culture environment, the induction success rate is low, and the cell state is difficult to control, and the drug resistance is unstable.
In addition, the drug-resistant cell lines induced by these two methods have obvious differences in cell morphology and biological characteristics, and belong to different cell sublines, resulting in the data obtained from drug screening in the two drug-resistant cell lines are often very different.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] This example is used to illustrate the culture of cisplatin-resistant lung cancer organoids.

[0053] (1) Preservation and transportation of cisplatin-resistant lung cancer tissue samples

[0054] Obtain bronchoscopic lung biopsy tissue from clinically confirmed cisplatin-resistant lung cancer patients as cisplatin-resistant lung cancer tissue samples. Cisplatin-resistant lung cancer tissue samples were placed in the In the preservation solution of RS-I Medium, it will be transported to the operating room within 48 hours of cold chain at 4 degrees.

[0055] (2) Acquisition of cisplatin-resistant lung cancer tissue cells

[0056] Place the cisplatin-resistant lung cancer tissue samples in a sterile petri dish, remove blood clots and necrotic tissue, rinse with sterile PBS for 3 times, and aspirate the PBS, then add 1.5 mg / mL type Ⅰ collagenase, use sterile The instrument cut the cisplatin-resistant lung cancer tissue sample into minced shape, and then added the enzym...

Embodiment 2

[0062] This example is used to illustrate the application of cisplatin-resistant lung cancer organoids in drug screening.

[0063] The cisplatin-resistant lung cancer organoid obtained in Example 1 was digested with TrypLE Express for 1 min, and then the hydrolyzed solution was collected and centrifuged at 400 rcf for 4 min. After centrifugation, the precipitate is collected, and a culture medium containing 10% low growth factor Matrigel is added to the collected precipitate to obtain a culture to be cultured. Aspirate the culture to be cultured above, and use 6×10 3 The inoculum amount per well is inoculated in a low-adsorption 96-well plate, and then the 96-well plate is placed in an incubator with 5% carbon dioxide at 37 ° C for 1 hour, and then 40 μL of culture medium is added to each well of the 96-well plate and continue to cultivate.

[0064] When the diameter of the culture in a single culture well is greater than 0.1 mm, and the culture confluence reaches 50%, add a...

Embodiment 3

[0066] The cisplatin-resistant lung cancer organoids were cultured according to the method of Example 1, except that the concentration of MMP-10 in the culture medium used in this example was 50 nM.

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Abstract

The disclosure relates to a method for culture of a cis-platinum drug-resistant lung cancer organoid. The method comprises the following steps: mixing cis-platinum drug-resistant lung cancer tissue cells with the medium and matrix gel to obtain a to-be-cultured substance, wherein the medium contains matrix metalloproteinase-10, and the concentration of the matrix metalloproteinase-10 is 50-200nM based on the medium; and carrying out culture amplification on the to-be-cultured substance to obtain the cis-platinum drug-resistant lung cancer organoid. According to the culture method for the cis-platinum drug-resistant lung cancer organoid provided by the invention, since the employed medium contains the matrix metalloproteinase-10 with specific concentration, and the matrix metalloproteinase-10 with the specific concentration can promote in-vitro growth of the cis-platinum drug-resistant lung cancer organoid, the success rate of the culture is high when the method provided by the disclosure is used for the culture of the cis-platinum drug-resistant lung cancer organoid.

Description

technical field [0001] The disclosure relates to the technical field of biomedicine, and in particular to a method for cultivating cisplatin-resistant lung cancer organoids, a medium for culturing cisplatin-resistant lung cancer organoids, cisplatin-resistant lung cancer organoids and their use in drugs. filter in the application. Background technique [0002] Cisplatin, also known as cis-dichlorodiammine platinum, is a widely used anti-cancer chemotherapy drug in clinical practice. It has broad-spectrum anti-tumor properties. It has a good therapeutic effect in the treatment of tumor diseases such as gastric cancer and bladder cancer. However, in the process of using cisplatin for chemotherapy, cisplatin resistance will occur, which will easily lead to the failure of chemotherapy, which greatly limits the use of cisplatin in tumor chemotherapy. [0003] Lung cancer is a common malignant tumor in the world, and its death toll accounts for about 18.4% of all cancer deaths. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2501/734C12N2513/00C12N2533/90G01N33/5011G01N2500/00
Inventor 孙志坚李程肖金平康平
Owner 中关村科技租赁股份有限公司
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