Nano Terra, Inc. proposes to develop novel 3D cellulosic scaffolds, constructed by stacking 2D layers, which enable the precise establishment of chemical gradients indicative of the in-vivo tumor microenvironment. Each layer is constructed using a simple guiding principle: minimize inhibition of phenotypic development by essentially ?staying out of the cells? way.? The scaffolds will contain extracellular matrix (ECM) mimics consisting of a minimal number of components, which can be stored in a dry state without refrigeration for periods of at least 6 months. In Phase I, in conjunction with the University of Maine and Seahorse Bioscience, we will demonstrate a scaffold that (1) supports the 3D growth of human tumor cells, including co-culture with multiple cell types;(2) replicates the in-vivo tumor microenvironment;(3) recapitulates the correct tumor phenotypes;(4) provides the ability to section 3D tumor constructs by simple destacking of layers;and (5) contains all reagents and chemical factors for tumor growth, without need for refrigeration. The Phase II effort will focus on scaffold validation, incorporating the scaffolds into commercial 24- and 96-well microplate formats, and manufacture of 5000 plates for commercial sale.