Establishment method of three-dimensional cell model

Abstract

The invention discloses an establishment method of a three-dimensional cell model. The method comprises the following steps: (1) inoculating cells in a culture-medium-containing culture dish, adding ferromagnetic nanoparticles into the culture medium, uniformly mixing, and culturing until the cell density reaches 80-90%, thereby obtaining ferromagnetic-nanoparticle-containing cells; and (2) reinoculating the ferromagnetic-nanoparticle-containing cells obtained in the step (1) in the culture-medium-containing culture dish, putting a magnetic control device which is jogged with the culture dish above the culture dish, and culturing for 18-36 hours, thereby obtaining the three-dimensional cell model, wherein the magnetic control device in the step (2) is jogged with the culture dish, and provided with a magnetic body. The method can quickly and efficiently establish the three-dimensional cell model, and the established three-dimensional cell model has the advantages of compact structure, low dispersion tendency and higher stability.

Description

technical field [0001] The invention relates to a method for constructing a cell culture model, in particular to a method for constructing a three-dimensional cell culture model. Background technique [0002] At present, most basic tumor-related research is based on two-dimensional cell culture, which is characterized by simple operation and easy study of phenotypic changes of single cells. However, such studies are difficult to simulate the tissue microenvironment of cells in vivo from the three-dimensional structural characteristics of tumor tissues, so they cannot accurately reflect the phenotypic characteristics of tumor cells in three-dimensional tissue. [0003] As a brand-new cell culture mode different from the traditional two-dimensional plate cell culture, three-dimensional cell culture can reflect the structural basis of the cell microenvironment in vitro to the greatest extent: 1) It can more truly reflect the process of tumor occurrence in vivo: cells Through t...

AI Technical Summary

Problems solved by technology

[0005] At present, the main problems in the construction of 3D cell models are as follows: (1) It is difficult to construct 3D cell models: firstly, it is necessary to construct dome-like matrigel clumps, and it is difficult to form dome-like matrigel after repeated freezing and thawing. The brand-new Matrigel also requires very skilled skills in shaping the dome shape, and once the dome shape is not well formed, such as eccentricity or collapse, it will directly lead to increased difficulty in the subsequent detection process, or even make it impossible to detect; if there is a three-dimensional If the matrigel culture model is in contact with the walls of the culture chamber, it will directly lead to the failure of model construction and subsequent experiments cannot be carried out

(2) It takes a long time to cultivate and form after construction: under normal circumstances, it takes about 7-14 days for tumor cells to form a spherical spatial structure on Matrigel

(3) Difficulty in detection: Conventional Matrigel is prone to swelling and softening after soaking in the culture medium for a week, so it is very easy to break the 3D Matrigel in the subsequent 3D immunofluorescent staining, making it impossible to complete subsequent staining and 3D reconstruction

[0006] The main reasons for the problems in the process of forming the existing 3D cell model are as follows: (a) The inhomogeneity of conventional manual methods: the traditional 3D model construction is closely related to the proficiency of the operator, but due to human factors It often causes the inhomogeneity of the three-dimensional culture model, which is manifested as: inhomogeneity in appearance (eccentricity, collapse) and inhomogeneity in the interior of the three-dimensional model (mainly caused by the flow of Matrigel during the shaping and solidification process)

(b) Shaping failure: Matrigel is prone to contact with the four walls of the culture chamber when shaping by conventional manual methods, and once contact occurs, due to the liquid tension, the Matrigel quickly flows to the walls of the culture chamber, forming a high-middle-low The "bowl-shaped" configuration, leading to shaping failure

This kind of situation is very easy to happen in the process of manual shaping, and it becomes one of the important reasons for the failure of shaping

(c) Difficulty in detection: Matrigel-based 3D cell culture models need to be soaked in culture medium for 7-14 days to grow. Matrigel is prone to swelling, softening and fragmentation after a week of immersion, which is very easy for subsequent 3D immunostaining There are problems such as model cracking and deformation, which affect the detection

Images