Development of a separable 3D cell co-culture system for the study of stem cell microenvironments

Stem cells represent an attractive cell choice for regenerative medicine applications due to their inherent ability to differentiate down multiple cell lineages given the correct cues. However, little is known concerning how the interactions, specifically the paracrine signaling effects, of native cells will influence the growth, proliferation, and differentiation of stem cells after implantation in vivo. This lack of fundamental knowledge necessitates the development of an in vitro model. Hydrogels are a biomaterial uniquely suited to the task of providing a tunable, cytocompatible environment in which to study these interactions between cell populations. By utilizing the technique of photolithography in conjunction with microfluidics, we developed and optimized a system to sequentially construct spatially segregated 3D co-culture constructs that will provide a basis for better understanding paracrine signaling between stem cells and other cell types native to orthopedic tissues. Additionally, by incorporating a degradable hydrogel interface between components, we demonstrated the separability of this system and the potential to isolate and recover individual populations after culture.Committee Member/Second Reader: Lu, Hang; Faculty Mentor: Temenoff, Johnn