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Host material for stereoscopic culture of breast cancer cells, preparation method and medical application

A technology of breast cancer cells and matrix materials, which is applied in the field of medical use for the prevention and treatment of breast cancer, can solve the problems of difficulty in forming a nutrient component/drug concentration gradient, lack of extracellular matrix components, and inability to effectively simulate, so as to enhance the effect of inhibiting tumors. , Efficient evaluation of drug efficacy, the effect of retaining dryness

Pending Publication Date: 2021-11-26
南京拓佳生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the limitations are also very obvious, such as: lack of extracellular matrix components, difficulty in forming a nutrient / drug concentration gradient, and the contact between cells is limited to one surface, which cannot effectively simulate the behavior of somatic cells, especially the drug resistance of cells sex

Method used

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  • Host material for stereoscopic culture of breast cancer cells, preparation method and medical application
  • Host material for stereoscopic culture of breast cancer cells, preparation method and medical application
  • Host material for stereoscopic culture of breast cancer cells, preparation method and medical application

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preparation example Construction

[0037] The preparation of the matrix material of the present invention involves a solution of component A: a solution of high-molecular biomaterials containing mercapto groups; a solution of component B: a solution of small-molecular biomaterials containing aldehyde groups.

[0038] The main component structural formulas of component A solution and component B solution are as follows image 3 and 4 shown

[0039] Component C solution is a solution containing an adhesion factor.

Embodiment 1

[0041] Matrix material preparation

[0042] Preparation of polymer material A containing mercapto saccharides (taking mercaptopropionic acid as an example):

[0043] Dissolve 0.05mmol carbohydrate polymer material in 10mL DMF solution, then add 0.001mmol DMAP and 0.1mmol EDCI to the system, stir in ice bath to room temperature for 0.5 hours, add 0.1mmol 3-mercaptopropionic acid to the system, and react After 3 hours, it was added to an ice-water bath, stirred for a while, and the aqueous layer was discarded. Use 0.05mol / L dilute hydrochloric acid solution to wash three times, and then use 0.05mol / L NaHCO to wash the residue 3 Wash three times. Finally, the residue was washed three times with double distilled water. The mercapto sugar polymer material A was obtained.

[0044] A preparation method for coupling a mercapto-aldehyde group into a matrix material, comprising the following steps: dissolving a mercapto-containing macromolecule biomaterial in a biocompatible solutio...

Embodiment 2

[0051] Matrix material for breast cancer cell culture

[0052] 1. Preparation of cell culture plate or cell culture dish suitable for breast cancer cell culture

[0053] Prepare component A solution, component B solution and component C solution according to the preparation scheme of the above-mentioned matrix material. Prepare a sterile cell culture plate or cell culture dish. This case uses a 24-well plate as an example.

[0054] 2. Spheroid growth and observation of breast cancer cells

[0055] The prepared matrix materials with different ratios were added to each well of a 24-well plate at 200 μL per well, and then placed in a 37°C incubator to continue incubating until the matrix was completely coagulated and formed. Add 300 μL of PBS to each well for rinsing in preparation for cell seeding.

[0056] Breast cancer cells or cells derived from breast cancer tissue (need to be processed into a single cell state in advance), and then placed in a cell culture medium contain...

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Abstract

The invention discloses a host material for stereoscopic culture of breast cancer cells, a preparation method and medical application. The host material for stereoscopic culture comprises a host material based on sulfydryl-aldehyde group coupling, and relates to precursor components: a solution A, a solution B and a solution C, wherein the solution A is a sulfydryl-containing polymer biological material solution; the solution B is an aldehyde-containing micromolecular biological material solution; and solution C is an adhesion factor-containing solution. The preparation method comprises the following steps: premixing A and C, standing, adding B, and uniformly mixing to form the host material. The invention provides a method for stereoscopic culture of breast cancer cells by using the host material, drug sensitivity analysis and other applications. After breast cancer cells are inoculated by using the host material, the cells are continuously cultured for several days to obtain a cell stereoscopic spherical structure, and the host material can be used for screening anti-cancer drugs, analyzing toxic and side effects and the like. According to the invention, the stereoscopic culture efficiency of the breast cancer cells is greatly improved, operation is easy, the detection period is shortened, and the method is suitable for stereoscopic culture and medical application research of the breast cancer cells.

Description

technical field [0001] The invention belongs to the field of medical application, and relates to a matrix material which can be used for three-dimensional culture of breast cancer cells, a preparation method thereof, and a medical application for preventing and treating breast cancer. Background technique [0002] At present, most cell culture methods are to grow cells on the surface of plastic culture dishes to form a monolayer of cells, which belongs to two-dimensional growth. In fact, in the body, cells do not simply exist in a single-layer structure, but adhere to each other and depend on each other, forming cells surrounded by other cells, tissues and extracellular matrix. The growth of cells in vivo is completely different from the culture environment of single-layer cells in vitro. Correspondingly, studying drugs based on two-dimensional growth methods cannot truly reflect the complex situation of drug application in vivo to a large extent. [0003] At present, most ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02C08L5/00C08K5/3477C08J3/24
CPCC12N5/0693C12N5/0631G01N33/5011C08J3/24C12N2533/30C12N2513/00C12N2503/02G01N2500/10C08J2305/00C08K5/3477
Inventor 王小明其他发明人请求不公开姓名
Owner 南京拓佳生物医药科技有限公司
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