Abstract
This paper describes an origami-inspired self-folding method to form three-dimensional (3D) microstructures of co-cultured cells. After a confluent monolayer of fibroblasts (NIH/3T3 cells) with loaded hepatocytes (HepG2 cells) was cultured onto two-dimensional (2D) microplates, degradation of the alginate sacrificial layer in the system by addition of alginate lyase triggered NIH/3T3 cells to self-fold the microplates around HepG2 cells, and then 3D cell co-culture microstructures were spontaneously formed. Using this method, we can create a large number of 3D cell co-culture microstructures swiftly with ease in the same time. We find that HepG2 cells confined in the 3D cell co-culture microstructures have an ability to enhance the secreted albumin compared to 2D system in a long culture period. The result indicates that the origami-based cell self-folding technique presented here is useful in regenerative medicine and the preclinical stage of drug development.
Highlights
A challenge for regenerative medicine and drug development is to fabricate in vitro 3D structures that mimic tissues in vivo
Our findings demonstrate that many 3D cell co-culture microstructures with NIH/3T3 cells covering HepG2 cells were rapidly with ease produced by the cell origami technique using an alginate sacrificial layer
We confirmed that the cells survived inside of the 3D microstructures and showed that the albumin secretion function of HepG2 cells co-cultured with NIH/3T3 cells inside of the 3D microstructures was increased in comparison with cells co-cultured on the microplates
Summary
A challenge for regenerative medicine and drug development is to fabricate in vitro 3D structures that mimic tissues in vivo. Several strategies have been developed to fabricate in vitro 3D cell-laden structures using a bottom-up technique[1,2,3,4,5,6], which involves micro-sized 3D cell-laden microstructures such as blocks[2], fibers[4,5,6] and spheroids[3,7] This approach allows one to control the size and shape of these microstructures, so that they can be handled and assembled to mimic in vivo tissue. We produced the 3D cell co-culture microstructures with fibroblasts (NIH/3T3) and hepatoma cells (HepG2) and rapidly using the cell origami technique This 3D cell co-culture microstructure provides www.nature.com/scientificreports/. We performed a viability assay and examined the hepatic function of the co-culture cells in the 3D microstructures by analysis of secreted albumin
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