CROSS-REFERENCE TO RELATED APPLICATION

This application is a National Stage of International Application No. PCT/KR2018/000652, filed on Jan. 15, 2018, which claims priority to Korean Patent Application No. 10-2017-0021375, filed on Feb. 17, 2017 in the Korean Intellectual Property Office, the disclosure of which is incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to transparent disc-shaped microparticles having polydopamine-modified surfaces and cell culture using the same.

BACKGROUND ART

The purpose of culturing animal cells is to produce various kinds of medical products including enzymes, hormones, vaccines, immunomodulators, anti-cancer drugs, exosomes, and cell therapeutic agents for treating diseases.

Animal cell culture method involves providing artificial in vivo-like environment for in vitro cell expansion. For some animal cells such as hybridoma or leukemia cell lines, scale-up expansion can be easily proceeded because they can be maintained by suspension culture. However, most of animal cells are adherent cells and they need surface to make attachment for in vitro expansion.

Since large number of stem/progenitor cells are needed for regenerative medicine and the related research, various scale-up manufacturing methods were developed to meet the need. Specifically, to increase cell yield per unit volume, area of cell-adherent surface was maximized by systems such as a microfiber, a hollow fiber, a ceramic matrix, or the like.

The microcarrier-based three-dimensional culture method was developed to overcome the limitations of conventional two-dimensional culture not only in volume efficiency but also in the way of handling. However, because the surface of microcarriers have three-dimensional contour, it is not easy to monitor the cell density or cell morphology.

The present invention provide an unique 3 dimensional culture method that can realize advantages of both the three-dimensional and two-dimensional culture by increasing surface efficiency and making two-dimensional cell observation possible at the same time.

A disc-shaped microparticle was prepared by photo-polymerizing a mixture of a photocurable synthetic polymer and linker molecules having a functional group that can be polymerized with photocurable substance and an alkoxysilyl group and the microparticle surface was coated with polydopamine to provide excellent cell adhesive properties and easy observation.

DISCLOSURE

Technical Problem

The present invention has been made in view of the above problems, and it is one object of the present invention to provide a microparticle having a two-dimensional plane formed by polymerization of a photocurable substance and a polydopamine-coated layer. The microparticles according to the present invention allow easy observation of cell morphology and density of attached cells while permitting three-dimensional culture.

Technical Solution

In accordance with one aspect of the present invention, provided is a disc-shaped microparticle for cell culture, including a polydopamine-coated surface layer, wherein the disc-shaped microparticle is formed by photocuring polymerization of a photocurable substance having an acrylate functional group and a linker having a functional group polymerizable with the photocurable substance and an alkoxysilyl group.

In accordance with another aspect of the present invention, provided is a method of preparing disc-shaped microparticles for cell culture having polydopamine-coated surfaces, the method including a) a step of preparing microparticles by photocuring polymerization of a mixture of a photocurable substance and a linker having a functional group polymerizable with the photocurable substance and an alkoxysilyl group; and (b) a step of adding a dopamine hydrochloride solution to the microparticles and stirring.

According to the present invention, the disc-shaped microparticle may include a coded polymeric microparticle core and a silica shell, wherein a carboxyl group or an amine group may be introduced to the surface of the silica shell.

According to the present invention, the disc-shaped microparticles may improve cell adhesive properties during cell culture, and may allow cell observation with an inverted microscope during cell culture, thus enabling easy observation.

According to the present invention, the stirring in the step of (b) may be performed for 40 to 80 minutes.

Advantageous Effects

When the disc-shaped microparticles coated with polydopamine according to the present invention are used in cell culture, cell adhesive properties are improved and an inverted microscope can be employed in a same way to observe cells in two-dimensional culture, which are major advantages when compared to conventional spherical microparticles.

