Culture medium, method and kit for rapidly culturing tumor organoids

A culture medium and organoid technology applied in the field of organoid culture to achieve the effect of improving the success rate and survival rate and shortening the required time

Active Publication Date: 2021-10-08
D1 MEDICAL TECH (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]Although a variety of tumor tissues can be successfully cultured into organoids in vitro using different methods and under different culture conditions, tumor org...

Method used

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  • Culture medium, method and kit for rapidly culturing tumor organoids
  • Culture medium, method and kit for rapidly culturing tumor organoids
  • Culture medium, method and kit for rapidly culturing tumor organoids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0184] Example 1 Intestinal cancer organoid culture

[0185] 1. Reagent preparation

[0186] 1) Rapid proliferation medium for tumor organoids: composed of medium components Advanced DMEM / F12, 20% sterile egg white extract and specific additive factors; the composition of specific additive factors: HEPES, 10mM; GlutaMAX, 2mM; 1XB27, 1:100(v / v); 1X N2, 1:100(v / v); N-acetylcysteine, 1mM; R-spondin-1, 500ng / ml; Noggin, 100ng / ml; EGF, 50ng / ml ; Nicotinamide, 10 mM; gastrin I, 10 nM; A83-01, 500 nM; ProstaglandinE2, 10 nM; SB202190, 5 μM; FGF basic, 50 ng / ml; Mycin B mixture (100 U / mL penicillin, 100 μg / ml streptomycin, 250 ng / mL amphotericin B), 1X; Y-27632dihydrochloride, 10 μM. The added factor was 4% of the total volume of the medium.

[0187] 2) Sterile egg white extract

[0188] The preparation method of sterile egg white extract is as follows: take 1-3 days of fresh eggs, separate egg whites and pass through 100 μm, 70 μm, and 40 μm cell strainers respectively; and then ...

Embodiment 2

[0210] Example 2 Culture of intestinal cancer organoids

[0211] 1. Reagent preparation

[0212] 1) Rapid proliferation medium for tumor organoids: It consists of the medium components Advanced DMEM / F12, 20% sterile egg white extract and specific additive factors; the composition of specific additive factors: HEPES, 10mM; GlutaMAX, 2mM; 1XB27, 1:100(v / v); 1X N2, 1:100(v / v); N-acetylcysteine, 1mM; R-spondin-1, 500ng / ml; Noggin, 100ng / ml; EGF, 50ng / ml ; Nicotinamide, 10 mM; gastrin I, 10 nM; A83-01, 500 nM; ProstaglandinE2, 10 nM; SB202190, 5 μM; FGF basic, 50 ng / ml; Mycin B mixture (100 U / mL penicillin, 100 μg / ml streptomycin, 250 ng / mL amphotericin B), 1X; Y-27632dihydrochloride, 10 μM. The added factor was 4% of the total volume of the medium.

[0213] The composition of sterile egg white extract, tissue preservation solution, tissue digestion solution, and cell mass collection solution is the same as that in Example 1.

[0214] 2. Culture method of intestinal cancer orga...

Embodiment 3

[0229] Example 3 Culture of liver cancer organoids

[0230] 1. Reagent preparation

[0231] 1) Rapid proliferation medium for tumor organoids: It consists of the medium components Advanced DMEM / F12, 30% sterile egg white extract and specific additive factors; the composition of specific additive factors: HEPES, 10mM; GlutaMAX, 2mM; 1XB27, 1:100(v / v); 1X N2, 1:100(v / v); N-acetylcysteine, 1mM; R-spondin-1, 500ng / ml; EGF, 50ng / ml; gastrin I, 10nM; A83-01, 500nM; Forskolin, 10μM; FGF basic, 50ng / ml; FGF4, 50ng / ml; FGF10, 100ng / ml; Streptomycin, 250ng / mL Amphotericin B), IX; Y-27632dihydrochloride, 10 μM. The added factor was 4% of the total volume of the medium.

[0232] The composition of sterile egg white extract, tissue preservation solution, tissue digestion solution, and cell mass collection solution is the same as that in Example 1.

[0233] 2. Culture method of liver cancer organoids

[0234] The present embodiment provides a method for culturing liver cancer organoids...

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Abstract

The invention relates to the field of organoid culture, in particular to a culture medium, a method and a kit for rapidly culturing tumor organoids. According to the culture medium for culturing the tumor organoids provided by the invention, the time for culturing the tumor organoids can be greatly shortened. The kit for rapidly culturing the tumor organoid comprises a culture medium for culturing the tumor organoid, a tissue preservation solution, a tissue digestion solution and a cell mass collection solution, and when the kit is used for culturing the tumor organoid, the success rate and survival rate of culturing the tumor organoid can be increased, an organoid model can be rapidly established within 2-5 days, which can be applied to subsequent experimental tests.

Description

technical field [0001] The present invention relates to the field of organoid culture, in particular, the present invention relates to a culture medium, a method and a kit for rapidly culturing tumor organoids. Background technique [0002] At present, the incidence and mortality of cancer are increasing year by year. In 2020, there will be 4.57 million new cancer cases and 3 million cancer deaths in China. The huge economic and social burden caused by tumors is very heavy. [0003] Surgery, radiotherapy, chemotherapy, targeted therapy, immunotherapy and endocrine therapy are the main means of tumor treatment. About 70% of these patients require radiotherapy and chemotherapy, but due to individual differences in patients, tumor drug resistance or radiation resistance is an important factor restricting the successful treatment of tumors. At present, the overall effective rate of clinical tumor drugs is low (about 30%), and unnecessary treatment and ineffective treatment of ...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00C12N2500/80
Inventor 富国祥李远闯林汉卿程春燕李俊强
Owner D1 MEDICAL TECH (SHANGHAI) CO LTD
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