Networked concave microwell arrays for constructing 3D cellspheroids

摘要

The engineered three-dimensional (3D) cell cultivation system for the production of multicellularspheroids has attracted considerable attention due to its improved in vivo relevance to cellularcommunications compared with the traditional two-dimensional (2D) cell culture platform. Theformation and maintenance of cell spheroids in a healthy condition is the critical factor for tissueengineering applications such as the repair of damaged tissues, the development of organreplacement parts and preclinical drug tests. However, culturing spheroids in conventional isolatedsingle wells shows limited yield and reduced maintenance periods due to the lack of proper suppliesof nutrition as well as intercellular chemical signaling. Here, we develop novel networked concavemicrowell arrays for the effective construction of 3D multi-cellular spheroids. The proposedmethod provides a suitable structure for the diffusion of oxygen, water-soluble nutrients andcytokines for cell–cell interactions between the spheroids in neighboring microwells. We havefurther demonstrated that hepatocyte spheroid cultured networked concave microwells showenhanced cell viability and albumin secretion compared to the un-networked control group overtwo weeks. Our results reveal that multi-cellular functionality can be tuned up by networkingindividual 3D spheroids withoutsupplying additional chemicals or biological supplements. Weanticipate our result to be useful in high-throughput cellular screening platforms to study cell–cellinteractions, in response to diverse chemical stimuli as well as the development of the in vivomimicking of the customized 3D tissue culture system.