Abstract
A cell culture on a scaffold has the advantages of functionality and easy handling, because the geometry of the cellular tissue is controlled by designing the scaffold. To create complex cellular tissue, scaffolds should be complex two-dimensional (2D) and three-dimensional (3D) structures. However, it is difficult to fabricate a scaffold with a 2D and 3D structure because the shape, size, and fabrication processes of a 2D structure in creating a cell layer, and a 3D structure containing cells, are different. In this research, we propose a micropatterning method for porous materials using the difference of the glass transition temperature between exposed and unexposed areas of a thick-photoresist. Since the proposed method does not require a vacuum, high temperature, or high voltage, it can be used for fabricating various structures with a wide range of scales, regardless of the materials used. Additionally, the patterning area can be fabricated accurately by photolithography. To evaluate the proposed method, a membrane integrated scaffold (MIS) with a 2D porous membrane and 3D porous material was fabricated. The MIS had a porous membrane with a pore size of 4 μm or less, which was impermeable to cells, and a porous material which was capable of containing cells. By seeding HUVECs and HeLa cells on each side of the MIS, the cellular tissue was formed with the designed geometry.
Highlights
An in vitro cell culture on a scaffold with microstructures can be used to create and elucidate biological functions and reproduction by constructing a biological structure similar to that which exists in vivo [1,2,3,4,5]
The micropatterning method for porous materials in this paper is composed of a partial reflow process dependent on the difference of the glass transition temperature between the exposed/unexposed areas of a thick-photoresist, and a releasing process using the sacrificial layer
The porous membrane patterned on the Alvetex scaffold was observed by SEM, and its size and fabrication error were calculated by image analysis
Summary
An in vitro cell culture on a scaffold with microstructures can be used to create and elucidate biological functions and reproduction by constructing a biological structure similar to that which exists in vivo [1,2,3,4,5]. Since the external environment and structure affect spontaneous tissue formation, the geometry of the cellular tissue can be controlled by designing the scaffold for the cell culture. The in vivo cellular tissue consists of a 2-dimensional (2D) boundary, such as barrier tissues, and a 3-dimensional (3D) dense cell structure, such as organs. To reproduce these tissues by cell seeding on the scaffold alone, the scaffold needs both 2D and 3D structures
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