Directed Self-Assembly of Large Scaffold-free Multicellular Honeycomb Structures

摘要

A significant challenge to the field of biofabrication is the rapid construction of large three dimensional (3D) living tissues and organs. Multi-cellular spheroids have been used as building blocks. In this paper, we create large multi-cellular honeycomb building blocks using directed self-assembly, whereby cell-to-cell adhesion, in the context of the shape and obstacles of a micromold, drives the formation of a 3D structure. Computer aided design, rapid prototyping and replica molding were used to fabricate honeycomb-shaped micro-molds. Nonadhesive hydrogels cast from these micro-molds were equilibrated in cell culture medium and seeded with two types of mammalian cells. The cells settled into the honeycomb recess, were unable to attach to the nonadhesive hydrogel and so cell-to-cell adhesion drove the self-assembly of a large multicellular honeycomb within 24 hours. Distinct morphological changes occurred to the honeycomb and its cells indicating the presence of significant cell-mediated tension. Unlike the spheroid, whose size is constrained by a critical diffusion distance needed to maintain cell viability, the overall size of the honeycomb is not limited. The rapid production of the honeycomb building unit, with its multiple rings of high density cells and open lumen spaces, offers interesting new possibilities for biofabrication strategies.