Composition, culture medium and method for 3D culture of laryngeal cancer tissue
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A composition and medium technology, applied in the field of 3D cultured laryngeal cancer tissue composition, can solve the problems of laryngeal cancer cell differences, laryngeal cancer tissue is difficult to culture into organoids, and laryngeal cancer basic and clinical research cannot be carried out, etc. achieve high consistency
Active Publication Date: 2022-05-06
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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Problems solved by technology
At present, ordinary culture techniques are mainly used for laryngeal cancer tissues. In the two-dimensional culture process, laryngeal cancer tissue cells are difficult or unable to fully express the pathophysiological characteristics of laryngeal cancer, so that the cultured primary laryngeal cancer cells and living laryngeal cancer The cells are quite different, so it is impossible to carry out basic and clinical research on laryngeal cancer
In 3D culture, it is difficult to culture laryngeal cancer tissues into organoids if a suitable medium cannot be found.
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Embodiment 1
[0037]The invention provides a culture medium for 3D culture of laryngeal cancer tissue, comprising: B27 serum-free additive, 0.5×; Wnt3A, 200ng / ml; N-acetylysteine, 1.5mM; EGF, 30ng / mL; Noggin, 150ng / mL; R-spondin 1, 200ng / mL; GSK1838705A, 2nM; FGF10, 40ng / mL; Nicotinamide, 20mM; Y-27632, 2μM; SB202190, 10μM; mL. The solvent was SAGM medium. The preparation method of the medium is as follows: formulating ingredients according to the final concentrations of all components, and then mixing with SAGM medium.
Embodiment 2
[0039] The invention provides a culture medium for 3D culture of laryngeal cancer tissue, comprising: B27 serum-free additive, 2×; Wnt3A, 50ng / ml; N-acetylysteine, 1mM; EGF, 100ng / mL; Noggin , 200ng / mL; R-spondin 1, 1000ng / mL; GSK1838705A, 2nM; FGF10, 10ng / mL; Nicotinamide, 5mM; Y-27632, 20μM; FGF2, 30ng / mL; SB202190, 10μM; . The solvent was SAGM medium. The preparation method of this culture medium is the same as that of Example 1.
Embodiment 3
[0041] The invention provides a culture medium for 3D culture of laryngeal cancer tissue, comprising: B27 serum-free additive, 1×; Wnt3A, 500ng / ml; N-acetylysteine, 0.15mM; EGF, 60ng / mL; Noggin, 50ng / mL; R-spondin 1, 500ng / mL; GSK1838705A, 0.5nM; FGF10, 50ng / mL; Nicotinamide, 10mM; Y-27632, 10μM; mL; trans-retinoic acid: 100 nM; BMP7: 10 ng / mL. The solvent was SAGM medium. The preparation method of this culture medium is the same as that of Example 1.
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Abstract
The invention provides a composition, a culture medium and a method for 3D culture of laryngeal cancer tissues. The culture medium comprises the following components according to final concentration: 0.5-2 * of a B27 serum-free additive; the concentration of Wnt3A is 50 to 500 ng/ml; the concentration of the N-acetylsteine is 0.15 to 1.5 mM; the concentration of the EGF is 20 to 100 ng/mL; the concentration of Noggin is 50 to 200 ng/mL; the concentration of the R-spondin 1 is 200 to 1000 ng/mL; the molecular weight of GSK1838705A is 0.2 nM to 2 nM; the concentration of the FGF10 is 10 to 50 ng/mL; 5 to 20 mM of Nicotinamide, and 10 to 20 mM of Nacotinamide; y-27632, 1 to 20 [mu] M; the solvent is an SAGM culture medium. According to the culture medium for the laryngeal cancer tissue organoid, disclosed by the invention, aiming at the culture and growth characteristics of laryngeal cancer tissue source cells, various cell factor components are selected and prepared according to a certain proportion, and laryngeal cancer cells can effectively form the organoid in the 3D culture medium; the formed organoid can maintain high consistency of tissue cell specificity, stem cell characteristics and genetic typing and high similarity of cellular morphology and physiological functions.
Description
technical field [0001] The invention relates to the technical field of organoid culture, in particular to a composition, a culture medium and a method for 3D culture of laryngeal cancer tissue. Background technique [0002] According to the latest 2018 Global Cancer Report released by the International Agency for Research on Cancer (IARC) under the World Health Organization, China ranks first in the world in terms of cancer incidence and mortality. Data shows that there are 3.804 million new cancer cases and 2.296 million cancer deaths in China, which is equivalent to more than 10,000 cancer diagnoses and more than 6,000 cancer deaths in my country every day. Cancer treatment is one of the biggest and most urgent problems in the biomedical field. On the one hand, there are more and more laboratory studies, on the other hand, the clinical transformation efficiency of new drugs is still low. Organoid culture provides a new technological platform for the rapid and effective d...
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