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Breast cancer organoid culture solution, preparation method and culture method

An organoid, breast cancer technology, applied in the field of biomedicine, can solve the problem of inability to effectively maintain the histopathological molecular typing characteristics of breast cancer, and achieve the effect of guiding clinical medication

Active Publication Date: 2022-06-07
杭州艾名医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a breast cancer organoid culture medium and its preparation method and culture method, so as to solve the problem that the existing breast cancer organoid culture medium cannot effectively maintain the pathological molecules of breast cancer tissue. The problem with categorical features

Method used

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  • Breast cancer organoid culture solution, preparation method and culture method
  • Breast cancer organoid culture solution, preparation method and culture method
  • Breast cancer organoid culture solution, preparation method and culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0031] An organoid culture medium for breast cancer, including basal culture medium Advanced DMEM / F-12 and L-propanyl-L-glutamine 2mM, green chain double antibody 100U / mL, N2 additive 1× (vol / vol), B27 additive 1× (vol / vol), N-acetyl-L-cysteine 0.09mM, SB 203580 0.3 μM, niacinamide 5mM, human recombinant protein R-Spondin 1 30ng / mL, Head protein 100 ng / mL, epidermal cell growth factor rhEGF 8 ng / mL, Neuregulin 3 5 nM, estradiol valerate 10 nM, Sovesudil 10 μM, HEPES 10 mM, and A83-01 0.5 μM.

[0032] The preparation method of the above-described breast cancer organoid culture solution, comprising the following steps: take the basal culture liquid AdvancedDMEM / F-12, first add L- alanyl-L- glutamine, N2 additive, B27 additive, N-acetyl-L-cysteine and niacinamide, mix evenly and wait for the solution to stabilize, and then add the remaining components, specifically: add blue chain double antibody, SB203580, growth factor and system stabilization factor, mix evenly, and prepare.

Embodiment 2

[0034]An organoid culture medium for breast cancer, including basal culture medium Advanced DMEM / F-12 and L-propanyl-L-glutamine 2mM, green chain double antibody 100U / mL, N2 additive 1× (vol / vol), B27 additive 1× (vol / vol), N-acetyl-L-cysteine 0.09mM, SB 203580 0.3 μM, niacinamide 5mM, human recombinant protein R-Spondin 1 30ng / mL, Head protein 100 ng / mL, epidermal cell growth factor rhEGF 8 ng / mL, Neuregulin 3 5 nM, estradiol valerate 10 nM, HEPES 10 mM, and A83-01 0.5 μM.

[0035] The preparation method of the above-described breast cancer organoid culture solution, comprising the following steps: take the basal culture liquid AdvancedDMEM / F-12, first add L- alanyl-L- glutamine, N2 additive, B27 additive, N-acetyl-L-cysteine and niacinamide, mix evenly and wait for the solution to stabilize, and then add the remaining components, specifically: add blue chain double antibody, SB203580, growth factor and system stabilization factor, mix evenly, and prepare.

Embodiment 3

[0037] An organoid culture medium for breast cancer, including basal culture medium Advanced DMEM / F-12 and L-propanyl-L-glutamine 2mM, green chain double antibody 100U / mL, N2 additive 1× (vol / vol), B27 additive 1× (vol / vol), N-acetyl-L-cysteine 0.09mM, SB 203580 0.3 μM, niacinamide 5mM, human recombinant protein R-Spondin 1 30ng / mL, Cephaloin 100 ng / mL, epidermal cell growth factor rhEGF 8 ng / mL, estradiol valerate 10 nM, Sovesudil 10 μM, HEPES 10 mM, and A83-01 0.5 μM.

[0038] The preparation method of the above-described breast cancer organoid culture solution, comprising the following steps: take the basal culture liquid AdvancedDMEM / F-12, first add L- alanyl-L- glutamine, N2 additive, B27 additive, N-acetyl-L-cysteine and niacinamide, mix evenly and wait for the solution to stabilize, and then add the remaining components, specifically: add blue chain double antibody, SB203580, growth factor and system stabilization factor, mix evenly, and prepare.

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Abstract

The invention discloses a breast cancer organoid culture solution as well as a preparation method and a culture method thereof. The culture solution is prepared from the following components: a basic culture medium, 1 to 5mM of L-alanyl-L-glutamine, 90 to 110U / mL of a blue chain double antibody, 0.5 to 1 percent (vol / vol) of an N2 additive, 1 to 2 percent (vol / vol) of a B27 additive, 0.06 to 0.1 mM of non-essential amino acid, 0.1 to 0.5 mu M of SB 203580, 3 to 8mM of nicotinamide, a growth factor and a system stabilizing factor. According to the culture solution disclosed by the invention, not only can very high cell and tissue activity be maintained in a culture process of breast cancer tissue cells, but also the breast cancer tissue cells can be cultured by simulating a disordered hormone environment in the body of a patient; the expression levels of the estrogen receptor PR and the progestational hormone receptor ER during preservation of the in-vitro cultured breast cancer organoid sample can better express the inherent pathological characteristics of the estrogen receptor PR and the progestational hormone receptor ER.

Description

Technical field [0001] The present invention relates to the field of biomedical technology, specifically to a breast cancer organoid culture solution and preparation method and culture method. Background [0002] Organoids are "micro-organoid-like" micro-organoid structures that are cultured under 3D conditions in vitro. Organoids have a wide range of uses, among which the cell clumps obtained by the isolation of tumor patients' tumor tissues can be tested for tumor susceptibility after the tumor organoids are constructed in vitro, and tumor organoids are used to replace patients with drug trials, and then guide clinical medication to achieve personalized and precise treatment of tumor patients. [0003] First, breast cancer is made up of several different molecular subtypes: Luminal A subtype (ER+ / PR+, HER2-), Luminal B subtype (ER+ / PR+, HER2+), HER2-positive (ER- / PR- / HER2+), and triple-negative (ER- / PR- / HER2-) breast cancer. There are clear differences in genomic variation betw...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/09
CPCC12N5/0631C12N5/0693C12N2500/46C12N2500/32C12N2501/10C12N2501/998C12N2501/11C12N2500/35C12N2501/80C12N2501/00C12N2500/30C12N2501/16
Inventor 邢华杨游明辉
Owner 杭州艾名医学科技有限公司
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