A kind of bladder cancer organoid culture medium and preparation method and application

A culture medium and organoid technology, which is applied in the field of bladder cancer organoid culture medium and preparation, can solve the problems of high cost and low success rate of bladder cancer organoids, and achieve the effects of high success rate, less drug consumption and short time

Active Publication Date: 2022-07-08
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method in this patent mainly uses the Transwell chamber, and the cost of this chamber is relatively high
[0009] In addition, other currently existing organoid preparation technologies have a low success rate in the preparation of bladder cancer organoids, and the aggrewell of the industrialized STEMCELL company hardly forms a ball during the preparation process.

Method used

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  • A kind of bladder cancer organoid culture medium and preparation method and application
  • A kind of bladder cancer organoid culture medium and preparation method and application
  • A kind of bladder cancer organoid culture medium and preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] A bladder cancer organoid culture medium is composed of the following components and dosages: cell culture medium 8mM, fibroblast growth factor 120ng / mL, hydrogen ion buffer 8mM, antibiotic 0.8%, epidermal growth factor 45ng / mL, recombinant protein 180ng / mL, Secreted Protein 460ng / mL, Human Gastrin 1 8nM, Nicotinamide or Nicotinamide 8mM, N-Acetyl Cysteine ​​1mM, Normocin 0.15%, Human Leukocyte Antigen 1.8%, Inhibitor 14.5uM and basement membrane matrix 36%; others are Advanced DMEM / F12 and trace solvent, Advanced DMEM / F12 is supplemented to the required volume of the medium. Trace amount of solvent, the solvent can be NAOH, sodium phosphate, Tris, sterile water, complete medium and DMSO. The complete medium is the medium of 90% DMEM+10% FBS+1% double antibody.

[0064]Among them, the component concentration is the final concentration of each component in the organoid medium, the measurement unit nM is the molar concentration, ng / mL is the mass concentration, and % is ...

Embodiment 2

[0074] Other contents are as in Example 1, a bladder cancer organoid culture medium is composed of the following components and dosages: cell culture medium 12mM, fibroblast growth factor 145ng / mL, hydrogen ion buffer 12mM, antibiotic 1.2%, epidermal cell growth Factor 55ng / mL, Recombinant protein 220ng / mL, Secreted protein 520ng / mL, Human Gastrin 1 12nM, Nicotinamide or Nicotinamide 12mM, N-acetylcysteine ​​1.5mM, Normocin 1.22 %, human leukocyte antigen 2.2%, inhibitor 16uM and basement membrane matrix 38.5%; others are Advanced DMEM / F12 and trace solvent, Advanced DMEM / F12 is supplemented to the required volume of the medium. Trace amount of solvent, the solvent can be NAOH, sodium phosphate, Tris, sterile water, complete medium and DMSO. In the experiment, the total amount of the general medium was 100 ml. The preparation method of the medium is a conventional technique.

Embodiment 3

[0076] Other contents are as in Example 1, a bladder cancer organoid culture medium is composed of the following components and dosages: cell culture medium 11mM, fibroblast growth factor 135ng / mL, hydrogen ion buffer 9mM, antibiotic 0.9%, epidermal cell growth Factor 48ng / mL, recombinant protein 190ng / mL, secreted protein 510ng / mL, human Gastrin 1 11nM, nicotinamide or nicotinamide 11mM, N-acetylcysteine ​​1.2mM, Normocin 1% , human leukocyte antigen 1.9%, inhibitor 15.5uM and basement membrane matrix 38%; others are Advanced DMEM / F12 and trace solvent, Advanced DMEM / F12 is supplemented to the required volume of the medium. Trace amount of solvent, the solvent can be NAOH, sodium phosphate, Tris, sterile water, complete medium and DMSO. In the experiment, the total amount of the general medium was 100 ml. The preparation method of the medium is a conventional technique.

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Abstract

The invention belongs to the field of tumor organoid culture, and in particular relates to a bladder cancer organoid culture medium, a preparation method and application thereof. The culture medium comprises the following components and dosages: a cell culture medium of 8-12 mM, a fibroblast growth factor of 120-120 mM 145ng / mL, hydrogen ion buffer 8‑12mM, antibiotic 0.8‑1.2%, epidermal growth factor 45‑55ng / mL, recombinant protein 180‑220ng / mL, secreted protein 460‑520ng / mL, human gastrin 8‑ 12nM et al. The method of the invention has good operability, high repeatability, rapidity, can be formed within 24 hours, the success rate is more than 90%, and the immune cells in the tumor are successfully retained, which can be used for the occurrence and development of bladder cancer, and the screening of tumor immune drugs. and in vitro susceptibility testing studies provide an ideal research model.

Description

technical field [0001] The invention belongs to the field of tumor organoid culture, and in particular relates to a bladder cancer organoid culture medium and a preparation method and application. Background technique [0002] Organoids are three-dimensional micro-organs that are highly similar to source tissues and organs and can be used in the laboratory from human stem cells (immature cells that differentiate into any cell type in the human body and play a key role in tissue function and regeneration). role) cultured, that is, taking some cells from the patient's body and culturing them in a petri dish, it can replicate some characteristics of the patient's tumor and be used to build a disease model. Organoids are essentially incarnations of a patient's tumor, capable of growing to about 1mm in diameter, resembling a patient's tumor in appearance, and sharing many of the same molecular and genetic features, which over time develop genetic changes, a phenomenon Known as c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N5/0684C12N2770/32321C12N2501/119C12N2501/115C12N2501/117C12N2501/11C12N2501/415C12N2501/155C12N2501/15C12N2501/998C12N2533/90C12N2501/727C12N2501/345C12N2500/38C12N2500/32C12N2500/60C12N2509/00C12N2509/10
Inventor 国世上龚芝伊
Owner WUHAN UNIV
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