NSF/FDA Scholar In Residence: 3D Cell Adhesion Assay for Cellularized Scaffold Characterization and Enhancement

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ABSTRACTPI: Fisher, JohnProposal Number: 16410873D Printing (3DP) is increasing tissue engineering (TE) applications through rapid manufacture of custom scaffolds wherein precision architectures can mimic native tissue. TE applications involving multipotent stem cells, 3DP constructs, and extracellular matrix (ECM) materials have become increasingly prevalent. In order to translate these technologies as therapeutics it is necessary to understand cell behavior within the 3D environment. An ongoing FDA-University of Maryland (UMD) collaboration has led to the development and characterization of a cell adhesion centrifugation (CAC) assay to enhance isolation and differentiation of mesenchymal stem cells (MSCs) from bone marrow. This proposal will enhance the utility of the established assay for use with 3D constructs. It will also provide useful insight into cell-scaffold interaction in three dimensions that will inform standardization criteria. The work will partner researchers, faculty and students at the University of Maryland and the FDA.The development of the CAC assay to accommodate 3D scaffold designs has the potential to minimize the translational distance for TE therapies by providing a mechanism to characterize and purify cells within 3D constructs. The cellular capture device will stabilize 3D scaffolds and allow characterization of non-adherent cells based on integrin-ligand binding. The key aims are: 1) investigate MSC behavior and purification from bone marrow seeded onto 3D constructs through adaptation of the CAC assay wherein the impact of centrifugation force, incubation time, and 3D construct architecture will be evaluated: 2) investigate the interaction of MSCs with 3D ECM-polymer scaffolds through examination of multi-lineage differentiation potential enabled by ECM additives to 3DP scaffold adhesion and differentiation capacity: and 3)investigate the capture and characterization of non-adhered cells collected using the CAC assays. This will allow the FDA to further develop the CAC assay and to evaluate its utility as a method to characterize the cellular component 3D construct based therapies.