InVERT molding for scalable control of tissue micro-architecture

Complex tissues contain multiple cell types that are hierarchically organized within morphologically and functionally distinct compartments. Construction of engineered tissues with optimized tissue architecture has been limited by tissue fabrication techniques, which do not enable versatile microscale organization of multiple cell types in tissues of size adequate for physiologic studies and tissue therapies. Here, we present an ‘Intaglio-Void/Embed-Relief Topographic (InVERT) molding’ method for microscale organization of many cell types, including induced pluripotent stem cell (iPS)-derived progeny, within a variety of synthetic and natural extracellular matrices and across tissues of sizes appropriate for in vitro, pre-clinical, and clinical biologic studies. We demonstrate that compartmental placement of non-parenchymal cells relative to primary or iPS-derived hepatocytes and hepatic compartment microstructure and cellular composition modulate hepatic functions. Configurations found to be optimal in vitro also result in superior survival and function after transplantation into mice, demonstrating the importance of architectural optimization prior to implantation. Two replicates of iPS-Hepatocytes cultured alone for 1 day or co-cultured with mouse J2 stromal cells for 1 day. One replicate of a mixture of iPS-Hepatocytes cultured alone for 1 day and then combined with J2 stromal cell RNA after RNA purification. Pure J2 stromal cell RNA was also hybridized to they Human chips but those hybridizations did not produce usable data and have not been included in this study.