3D printed synthetic tissues for patterned interactions with cellular populations

Cellular communities, consisting of cells (microbial and/or eukaryotic) living and interacting in various environments, are starting to be used in applications ranging from environmental remediation, agriculture, food science, bioproduction, and biomedicine. Moreover, it is increasingly being realized that communities of interacting cells underpin many aspects of human health (i.e. microbiomes). A global research priority therefore is to understand and engineer these communities for our own goals. Patterned population gene expression in cellular communities is critical for the establishment and development of both microbial communities and eukaryotic tissues. However, external control over target-cell populations is hugely limited due to the lack of smart-patterned release systems that can integrate and deliver effector molecules to cells when required. Here we propose to solve this, by using a custom-built 3D printer to build a smart-patterned release system for controlling population gene expression in cells with high spatial and temporal resolution. These printed systems will comprise of 100s of pL-sized aqueous droplets networked by interfaced lipid bilayers, of which we call synthetic tissues. Critically, we will develop these synthetic tissues to function in aqueous environments where encapsulated effector molecules will be released through membrane proteins present in the connected bilayers. Further, we will develop these 3D printed patterned release systems to be robust and adaptive to their external environment, and validate our system by interrogating patterned gene expression in both defined bacterial and mammalian cell populations.