Pleuroperitoneal fluid type organ culture medium and method and drug sensitive testing method
A culture method and culture medium technology, applied in the field of organoid culture, can solve the problems of non-cancerous pleural effusion organoid culture methods, differences, operability and good repeatability, and achieve the maintenance of tissue cell specificity and mild effect Effect
Active Publication Date: 2019-08-16
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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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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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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The invention discloses a pleuroperitoneal fluid type organ culture medium and method and a drug sensitive testing method. The pleuroperitoneal fluid type organ culture medium can effective maintain specificity of tissue cells and characteristics of stem cells and ensure high consistency of genetic typing as well as high similarity of tissue shape. Besides meeting the demands of scientific research, the pleuroperitoneal fluid type organ culture medium can be applied to the aspect of clinical medication guidance, and through in vitro organ culture, provide patients with a beneficial option on 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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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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