Culture medium and culture method for colorectal cancer organoids

A culture medium and organoid technology, applied in the field of biomedicine, can solve the problems of less research and reports on colon cancer organoid culture methods, test procedures, operation steps, culture conditions, and medium formulations, etc. Achieve the effect of reducing the risk of microbial contamination, reducing cytotoxicity and inhibitors, and improving success and survival rates

Active Publication Date: 2020-07-10
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although a variety of tumor tissues can be successfully cultured into organoids in vitro using different methods and under different culture conditions, there are currently few studies and reports on the culture methods of intestinal cancer organoids, especially the specific test procedures and operating steps , culture conditions, that is, there are not many reports on the medium formula

Method used

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  • Culture medium and culture method for colorectal cancer organoids
  • Culture medium and culture method for colorectal cancer organoids
  • Culture medium and culture method for colorectal cancer organoids

Examples

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Embodiment 1

[0038] This embodiment provides a culture medium for intestinal cancer organoids, including basal medium Advanced DMEM / F12, specific additive factors and sterile water; wherein, the basal medium Advanced DMEM / F12 and the sterile water The mass ratio is: 99:1; the specific addition factors include: vitamin A-free B27, 1×; N-acetylcysteine, 1.5 μM; EGF, 50ng / ml; Noggin, 200ng / ml; R-spondin 1, 200ng / ml; Wnt3a, 100ng / ml; CHIR99021, 5μM; thiazovivin, 2μM; Gastrin I, 15ng / ml; penicillin and streptomycin mixture, 1.2×; The concentration is based on the concentration in the mixture of basal medium and sterile water.

Embodiment 2

[0040]This embodiment provides a culture medium for intestinal cancer organoids, including basal medium Advanced DMEM / F12, specific additive factors and sterile water; wherein, the basal medium Advanced DMEM / F12 and the sterile water The mass ratio is: 99:1; the specific addition factors include: B27 without vitamin A, 2×; N-acetylcysteine, 1 μM; EGF, 80ng / ml; Noggin, 100ng / ml; R-spondin 1, 400ng / ml; Wnt3a, 80ng / ml; CHIR99021, 2μM; thiazovivin, 4μM; Gastrin I, 10ng / ml; penicillin-streptomycin mixed solution, 1×; The concentration is based on the concentration in the mixture of basal medium and sterile water.

Embodiment 3

[0042] This embodiment provides a culture medium for intestinal cancer organoids, including basal medium Advanced DMEM / F12, specific additive factors and sterile water; wherein, the basal medium Advanced DMEM / F12 and the sterile water The mass ratio is: 99:1; the specific addition factors include: B27 without vitamin A, 2×; N-acetylcysteine, 2μM; EGF, 20ng / ml; Noggin, 400ng / ml; R-spondin 1, 600ng / ml; Wnt3a, 150ng / ml; CHIR99021, 5μM; thiazovivin, 2μM; Gastrin I, 20ng / ml; penicillin-streptomycin mixture, 1.2×; All are subject to its concentration in the mixture of basal medium and sterile water.

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Abstract

The present invention provides a culture medium and a culture method for colorectal cancer organoids. The culture medium comprises a basic culture medium Advanced DMEM/F12, specific additive factors and sterile water; wherein a mass ratio of the basic culture medium Advanced DMEM/F12 to the sterile water is 99:1; and the specific additive factors comprise B27 without vitamin A, N-acetylcysteine, EGF, Noggin, R-spondin 1, Wnt3a, CHIR99021, thiazovivin, Gastrin I, a penicillin-streptomycin mixture and Primocin. The culture medium is used to culture the colorectal cancer organoids, can maintain morphological structures and genetic characteristics of primary tissues, effectively reduces risks of microbial contamination in colorectal cancer culture, and improves success rate and survival rate of colorectal cancer organoid culture.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, specifically relates to a culture medium and its application, and more specifically relates to a culture medium and a culture method of intestinal cancer organoids. Background technique [0002] Colon cancer is one of the top ten malignant tumors in my country, with an incidence rate of about 24 / 100,000, and it is also one of the tumors with the fastest-rising incidence rate in the past 20 to 30 years. Bowel cancer includes colon cancer and rectal cancer. The incidence of bowel cancer from high to low is rectum, sigmoid colon, cecum, ascending colon, descending colon, and transverse colon. In recent years, there is a trend of developing to the proximal end (right colon). In the early stages of colon cancer, there are no symptoms, or the symptoms are not obvious, only discomfort, indigestion, and occult blood in the stool. As the cancer develops, symptoms gradually appear, manifested as chang...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/32C12N2501/11C12N2501/345C12N2501/405C12N2501/415C12N2509/00
Inventor 黄敏陈泽新廖传荣
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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