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A kind of culture medium and culture method of gastric cancer organoids

A culture medium and organoid technology, applied in the field of biomedicine, can solve the problems of few researches and reports on gastric cancer tumor organoid culture methods, and few reports on organoid culture medium formula, so as to reduce the risk of microbial contamination and reduce the risk of cell Toxicity and Inhibitors, Effects to Increase Success and Survival

Active Publication Date: 2020-11-03
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 few studies and reports on the culture methods of gastric cancer tumor organoids, especially the formula of organoid culture medium is still unknown. Not many reports

Method used

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  • A kind of culture medium and culture method of gastric cancer organoids
  • A kind of culture medium and culture method of gastric cancer organoids
  • A kind of culture medium and culture method of gastric cancer organoids

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Embodiment 1

[0038] This embodiment provides a culture medium for gastric cancer organoids, including basal medium 1640, specific additive factors and sterile water; wherein, the mass ratio of the basal medium 1640 to the sterile water is: 99:1 The specific addition factors include: vitamin A-free B27, 1×; N-acetylcysteine, 2 μM; EGF, 50ng / ml; Noggin, 200ng / ml; R-spondin 1, 200ng / ml; Wnt3a, 100ng / ml ml; CHIR99021, 5μM; thiazovivin, 2μM; Gastrin I, 20ng / ml; The concentration of each component of the additive factor is based on its concentration in the mixture of basal medium 1640 and sterile water.

Embodiment 2

[0040] This embodiment provides a culture medium for gastric cancer organoids, including basal medium 1640, specific additive factors and sterile water; wherein, the mass ratio of the basal medium 1640 to the sterile water is: 99:1 The specific addition factors include: vitamin A-free B27, 2×; N-acetylcysteine, 1 μM; EGF, 80ng / ml; Noggin, 150ng / ml; R-spondin 1, 400ng / ml; Wnt3a, 150ng / ml ml; CHIR99021, 5μM; thiazovivin, 2μM; Gastrin I, 25ng / ml; The concentration of each component of the additive factor is based on its concentration in the mixture of basal medium and sterile water.

Embodiment 3

[0042]This embodiment provides a culture medium for gastric cancer organoids, including basal medium 1640, specific additive factors and sterile water; wherein, the mass ratio of the basal medium 1640 to the sterile water is: 99:1 The specific addition factors include: vitamin A-free B27, 1.5×; N-acetylcysteine, 4.5 μM; EGF, 80ng / ml; Noggin, 250ng / ml; R-spondin 1, 150ng / ml; Wnt3a, 150ng / ml; CHIR99021, 2.5μM; thiazovivin, 2.5μM; Gastrin I, 40ng / ml; Valproic acid, 1.5mM; penicillin-streptomycin mixed solution, 2×; mg / ml; the concentration of each component of the above-mentioned specific addition factor is based on its concentration in the mixture of basal medium and sterile water.

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Abstract

The invention provides a culture medium for stomach cancer organs and a culture method. The culture medium comprises a basic culture medium 1640, specific addition factors and sterile water, wherein the mass ratio of the basic culture medium 1640 to the sterile water is 99:1; and the specific addition factors comprise B27 without vitamin A, N-acetylcysteine, EGFs (epidermal growth factors), Noggin, R-spondin 1, Wnt3a, CHIR99021, thiazovivin, Gastrin I, valproic acid, a penicillin and streptomycin mixed liquid, amphotericin B and Primocin. The culture medium can be adopted to culture the stomach cancer organs, morphology structures and gene characteristics of primary tissue can be maintained, the risk of microorganism pollution in stomach cancer culture can be effectively reduced, and the success rate and the survival rate of stomach cancer organ culture can be increased.

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 gastric cancer organoids. Background technique [0002] Gastric cancer is a malignant tumor originating from the gastric mucosal epithelium, and it is one of the common malignant tumors in my country. Gastric cancer can occur in any part of the stomach, and more than half of them occur in the gastric antrum. The greater curvature, the lesser curvature, and the anterior and posterior walls can be involved. The vast majority of gastric cancer is adenocarcinoma. There are no obvious symptoms in the early stage, or non-specific symptoms such as epigastric discomfort and belching, which are often similar to the symptoms of chronic gastric diseases such as gastritis and gastric ulcer, and are easily ignored. Therefore, the early diagnosis of gastric cancer in my co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/30C12N2500/32C12N2501/11C12N2501/345C12N2501/405C12N2501/415C12N2509/00C12N2509/10
Inventor 王哲君李刚徐丛
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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