Title: Two-photon fabricated scaffolds for controlled three-dimensional cell migration studies
Description: Two photon polymerization (TPP) is a fabrication technique that has been extensively used for making three-dimensional structures of different shapes and sizes with sub-micrometer resolution; but applications using this technique in biology are only numbered. Cell migration studies have progressed from two-dimensions (2D) to three-dimensions (3D) owing to the striking phenomenal differences observed between the two. This dissertation is aimed at bringing the two fields together to understand cellular phenomena in a more systematic and controlled way. This dissertation describes a novel methodology to study 3D cell migration. TPP was used to fabricate 3D interconnected scaffolds with precise architectural control at micrometer resolution. A live imaging and analysis system was developed to study cell migration within those scaffolds. Experiments were conducted to compare 2D and 3D cell migration and study the effects of matrix architecture on 3D cell migration for normal and tumor cell lines. Cell migration was quantified in terms of various cell motility parameters like cell speed, persistence and probability of motion. This system provided a platform for doing a controlled study on cell phenomena by varying any one parameter of the matrix independently without affecting any other parameter. Further, since the past decade, dendritic cell (DC) based immune therapy approaches have been developed for cancer therapeutics. However, most of the limitations of these approaches suggest that the DCs need to be improved in the quality of their maturation state and their migratory abilities to home towards the lymph node to start an immune response. The microfabrication technology was extended to do a controlled study for directed migration of DCs in the presence of a lymph-node chemokine.

A live migration assay to study chemotaxis was developed, which is much more informative than a lot of other chemotactic assays (e.g. transwell systems), that can only be used as endpoint assays. The microfabricated system was developed to study the effect of different chemokines and adjuvants on DC migration. Effect of the interplay between two different chemokines on cell migration was studied. The system was extended to study the effects of architecture and chemotaxis on dendritic cell migration by using a combination of fabrication techniques and controlled release strategies. A theoretical model for our system was also developed to simulate and explain the experimental results. These simulations could help us optimize the concentration and release profile of chemokines to result in more effective directed migration of cells. It could also be used to predict a range of scaffold architectural parameters for either promoting or restraining the motion of cells. These predictions could have implications in the development of more effective cancer vaccines.