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A kind of pleural and ascites organoid culture medium, culture method and drug susceptibility testing method

A culture method and organoid technology, which can be used in the field of medium for 3D culture of residual cancer tissue or cancer cells in pleural and ascites fluids of tumor patients after surgery, which can solve the problem of organoid culture methods, differences, operability, and differences in cancer-free pleural and ascites fluids. Good repeatability and other problems, to achieve the effect of maintaining tissue cell specificity and mild effect

Active Publication Date: 2021-02-09
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Under two-dimensional culture conditions, it is difficult or impossible for cancer cells to fully express the characteristics of their source cancer tissue, which makes the response of cultured cancer cells and their source cancer tissue different to drugs, which cannot be used as adjuvant drug guidance
[0005] Although a variety of human-derived tissues can be successfully cultured into organoids in vitro under different culture conditions, there are currently no studies and reports on the culture methods of cancerous pleural effusion organoids, especially the specific culture conditions, that is, the collection of cancer cells. Preservation method, medium for 3D culture, and drug susceptibility screening program as an auxiliary guide for postoperative medication have not been tried or reported yet. The patented formula of the present invention is moderately priced and has good operability and repeatability

Method used

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  • A kind of pleural and ascites organoid culture medium, culture method and drug susceptibility testing method
  • A kind of pleural and ascites organoid culture medium, culture method and drug susceptibility testing method
  • A kind of pleural and ascites organoid culture medium, culture method and drug susceptibility testing method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A culture medium component of pleural fluid organoids, the content of each component is as follows: B27, 50X dilution; N-acetylsteine, 2.5mM; EGF, 10ng / ml; Noggin, 100ng / ml; R-spondin 1, 500ng / ml or 30 % Conditioned medium; A83-01, 500nM; FGF10, FGF2, 25ng / ml; Nicotinamide, 10mM; Y-27632, 10μm; Prostaglandin E2, 1μM; SB202190, 10μm;

Embodiment 2

[0045] A preservation solution for pleural and ascites, the content of each component is as follows: B27 (0.5x), Hepes (5mM), dispase II (0.1%), EGF (10ng / ml), N2 (0.5x), Nicotinamide (5mM), Y- 27632 (5 μM), A83-01 (0.5 μM) and SB202190 (5 μM).

Embodiment 3

[0047] After the organoids are cultured, the composition of the digestive juice for collecting and digesting the organoids is DMEM / F12 containing 0.1% trypsin, 0.2% dispaseⅡ, 0.1% collagenaseⅢ, insulin 2.5μg / ml, Hepes0.5M, EGF 10ng / ml.

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Abstract

The invention discloses a pleural effusion organoid culture medium, a culture method and a drug sensitivity testing method. The culture medium of the invention can effectively maintain the high consistency of tissue cell specificity, stem cell characteristics, and genotyping, and the tissue morphology is also highly similar . In addition to meeting the needs of scientific research, in terms of clinical medication guidance, in vitro organoid culture provides a good and beneficial choice for patients' drug medication guidance.

Description

technical field [0001] The present invention relates to the field of organoid culture, and further relates to the field of pleural and ascites organoid culture, in particular to a culture medium for 3D culture of residual cancer tissue or cancer cells in the pleural and ascites of tumor patients, a method for culturing organoids and a method for drug sensitivity testing. Background technique [0002] Cancerous pleural effusion, also known as malignant pleural effusion, is one of the common complications of advanced cancer, and it is also the main clinical symptom or sign of some patients. Severe pleural and ascites can even be life-threatening. The diseases of malignant pleural effusion are common in lung cancer and breast cancer, followed by malignant lymphoma, ovarian cancer, malignant pleural mesothelial carcinoma, esophageal cancer, gastric cancer, cardia cancer and malignant tumors of unknown etiology. [0003] Surgery is currently the mainstay of treatment for many typ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02A01N1/02
CPCA01N1/0226C12N5/0693C12N2503/02G01N33/5011G01N2500/10
Inventor 郑鸿平邱培陈泽新
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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