Origami-based self-folding of co-cultured NIH/3T3 and HepG2 cells into 3D microstructures

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.

Advanced Bottom‐Up Engineering of Living Architectures

“…I,J) Self‐folding origami‐inspired co‐culture structures assembled from cell sheets. I,J) Reproduced under the terms of the CC‐BY Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/) . Copyright 2018, The Authors, published by Springer Nature.…”

“…Moreover, by controlling cell orientation in the 2D surface template, they could fabricate tubules with circumferentially and longitudinally oriented SMCs, thus mimicking the anisotropy seen in native tunica media and adventitia. Aiming to achieve more complex architectures, self‐folding co‐cultured cell sheets were obtained by culturing cells in origami‐inspired micromolded alginate substrates that release cell assemblies upon enzymatic degradation with alginate lyase . The resulting dodecahedron microstructures give rise to 3D co‐culture cell‐rich assemblies via a self‐folding process mediated by cell–cell traction force (Figure I,J).…”

Origami Biosystems: 3D Assembly Methods for Biomedical Applications

“…Strain engineering based on self‐rolling methods has been used to create a variety of 3D curved tissue scaffolds. These include single and multilayered rolls (Figure i,j) and vascular mimics . They highlight the advantages of origami approaches such as facile layering of different cells and matrix as is needed in several tissues including blood vessels, the ability to leverage state of the art 2D patterning techniques such as photopatterning, contact printing and soft‐lithography, and the ability for high‐throughput fabrication of curved and folded cellular geometries that can be hard to access by other methods.…”

Morphodynamic Tissues via Integrated Programmable Shape Memory Actuators

“…Origami is the art of folding paper into 3D shapes, and applying this strategy toward tissue engineering could create uniquely complex biological architectures . This strategy could integrate two broad tissue engineering paradigms: cell‐driven approaches in which the cells themselves direct and organize the tissue architecture; and externally driven strategies in which tissues are physically manipulated into desired shapes .…”

“…These techniques present limited options in terms of controlling the timing or reversibility of 3D shape actuation. More complex alternatives have also been proposed to change the shape of tissues including, leveraging the contractile activity of cells themselves by positioning cells at hinges between rigid plates, culturing contractile cells on anisotropically flexible substrates, or through cellular contraction of 3D collagen biomaterials . Unfortunately, these strategies are not broadly applicable, as they exclude tissues that do not incorporate mechanically contractile cells, require certain cell and tissue spatial arrangements, and cannot be triggered on‐demand to effect shape change at the appropriate time.…”