3D cell culture medium and preparation method thereof

Abstract

The invention discloses a 3D cell culture medium and a preparation method thereof. The preparation method includes: mixing oxidized alginic acid with sodium alginate in proportion, and utilizing distilled water to prepare a mixed sodium alginate solution; performing filtering in a sterile environment, then adding a concentrated culture medium and a serum / growth factor sequentially, and mixing thesubstances uniformly to obtain a culture solution; and finally, conducting mixing and crosslinking with a sterile calcium chloride solution to obtain the 3D cell culture medium. The 3D cell culture medium prepared by the method provided by the invention solves the problems that a sodium alginate solution can hardly achieve effective degerming and oxidized sodium alginate has a too low crosslinkingdegree due to a too high oxidation degree. Meanwhile, the plasticity of the culture medium is improved by adjusting the ratio of sodium alginate to oxidized sodium alginate, and the culture medium ismore suitable for proliferation and growth of cells. If cells need to be cultured, the cells needing culture are added into a culture solution according to certain inoculation concentration, the substances are mixed gently and uniformly, and finally the product is crosslinked with sterile calcium chloride so as to culture the cells.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and relates to a preparation method of a 3D cell culture substrate and the 3D cell culture substrate prepared by the method. Background technique [0002] Sodium alginate (SA), as a natural polymer material, has good biocompatibility. SA hydrogel has biological properties similar to extracellular matrix and is suitable for the construction of cell culture microenvironment. However, sodium alginate (SA) solution is highly viscous and difficult to sterilize by filtration. However, the sterilization methods for SA in the current literature usually use high temperature and high pressure sterilization, high temperature dry heat sterilization, ultraviolet sterilization and radiation sterilization. However, high temperature, high pressure and high temperature dry heat sterilization will destroy the formability of the hydrogel, while ultraviolet sterilization has the problem of poor sterili...

AI Technical Summary

Problems solved by technology

However, high temperature, high pressure and high temperature dry heat sterilization will destroy the formability of the hydrogel, while ultraviolet sterilization has the problem of poor sterilization effect, and radiation sterilization also has a great impact on the material properties of the hydrogel.

This greatly limits the scope of application of sodium alginate hydrogel as a 3D cell culture substrate in scientific research and clinical practice.

Due to the sterilization problem of SA, so far no SA3D culture cell matrix can be used for long-term cell culture

Images