Abstract
Collagen-based hydrogels are widely used for three-dimensional (3D) culture of mammalian cells because of their high cell-activating characteristis. However, techniques for preparing of cell-embedding collagen hydrogels with micrometer-size precision in perfusable, microfluidic devices have not been fully developed. In this study, we propose a facile strategy enabling microfabrication of collagen hydrogels in microfluidic devices. We used phosphate particle-embedding polydimethylsiloxane (PP-PDMS) as a gelation agent, which neutralizes the acidic collagen soltuion. The collagen solution near the surface of the PP-PDMS is selectively gelled. We fabricated micropatterns and tubular structures made of collagen hydrogel, both of which were used for perfusion culture of mammalian cells encapsulated in the hydrogel matrix. The presented approach would be applicable to various types of cell culture experiments.
Highlights
In recent years, researchers have developed perfusion culture devices of mammalian cells, which can reproduce the blood circulation system in the organs or tissues in our body
We fabricated two types of microfluidic devices partially made of phosphate particle-embedding polydimethylsiloxane (PP-PDMS); (1) flat PDMS channels equipped with phosphate particle (PP)-PDMS microposts for hydrogel patterning, and (2) channels entirely composed of PP-PDMS for creating tubular collagen hydrogels
3.1 Observation of gelation on the PP-PDMS plate First, we investigated if the collagen hydrogels were formed on the surface of the PP-PDMS because of the interaction with the embedded phosphate particles
Summary
Researchers have developed perfusion culture devices of mammalian cells, which can reproduce the blood circulation system in the organs or tissues in our body. Cells are surrounded by extracellular matrix (ECM) components, including collagen, laminin, fibronectin, and so on The use of these ECM-based hydrogels was proven to be significantly effective for culturing specific cell types, as represented by hepatocytes. When an acidic collagen solution was dropped onto or contacted with the surface of the PP-PDMS, the phosphate particles were dissolved, resulting in the partial neutralization of the collagen solution and the hydrogel formation at the vicinity of the PP-PDMS surface. We created tubular collagen structures in channels entirely composed of PP-PDMS (entirely PP-PDMS channel) Both of these structures were used to perform perfusion culture of mammalian cells embedded in the collagen hydrogel matrix, to investigate the usefulness of our approach for cell culture applications
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