Preparation method of double-layer cell ball

Abstract

The invention relates to a preparation method of a double-layer cell ball. The invention aims at providing a preparation method of the double-layer cell ball, comprehensively and truly reflecting the survival environment of in-vivo cells, and visually reproducing the interaction between two kinds of cells under certain conditions. The technical schemes for achieving the aims are as follows: the preparation method of the double-layer cell ball comprises the steps of: a. separately culturing two different kinds of cells; b. preparing a single cell suspension from one kind of the cells; c. controlling the single cell concentration in a bottle at 1*10<5>-1*10<6> cells / ml; d. standing the single cell suspension in a 37 DEG C incubator containing 5% of CO2 for 1-4 hours; e. carrying out rotation culture on the single cell suspension for rolling the single cell suspension into balls, and changing the single cell suspension once every 10-14 hours; and f. preparing a single cell suspension of the other kind of cells, and adding the single cell suspension to the culture liquid containing the cell balls obtained in step e, continuing rotation culture so that the two kinds of cells in a tube form a spherical cell mass with a clear boundary and a double-layer structure, and changing the suspension once every 10-14 hours. The preparation method of the double-layer cell ball provided by the invention is mainly used for three-dimensional culture of cells.

Description

technical field [0001] The invention relates to a method for making double-layer cell spheres, which is mainly suitable for three-dimensional culture of cells. Background technique [0002] At present, the traditional culture of adherent cells is a two-dimensional culture model, a model of cell monolayer culture, which lacks the support of the microenvironment in vivo, and the experimental results obtained cannot be consistent with the actual situation in vivo. Since the parenchymal cells are in close contact with the surrounding mesenchymal cells in the body, there is an interaction between them. The normal interaction can maintain the homeostasis and the normal operation of the body, while the abnormal interaction can lead to diseases, such as tumors. Studies have found that the occurrence of tumors is not only caused by tumor cells, its local microenvironment also plays a vital role, and even determines the fate of tumors. Tumor cells are always in close contact with the...

AI Technical Summary

Problems solved by technology

Tumor cells are always in close contact with the surrounding mesenchymal cells during the development process. When the effect factors in the local microenvironment reach a certain concentration, the interaction between the two can lead to a series of structural, functional and metabolic changes in tumor cells. Therefore, this change cannot be observed by ordinary cell culture methods due to the limited number of cells and the dilution of the culture medium. At present, there are some three-dimensional culture models, but they all rely on the effect of some kind of gel, such as hydrogel, collagen gel, etc. Glue is constructed and cannot truly reflect the living environment of cells in the body