Formation of Embryoid Bodies and Encapsulation of Mouse Embryonic Stem Cells via Stereolithography

Embryonic stem cells have the ability to differentiate into cells of the three germ layers – ectoderm, mesoderm and endoderm. Embryonic stem cells can be very promising in developing especially for tissue engineering and regenerative medicine thereby could replace organ donations which are very limited and expensive. Mouse embryonic stem cells (mESCs) will be formed into embryoid bodies (EBs) or sphere of cells composed of specific cell concentrations. As differentiation of stem cells is a density dependent phenomenon, we will first form EBs of different cell densities to characterize their packing efficiency and density. Finally, individual mESCs will also be encapsulated in hydrogels using stereolithography for long-term differentiation. Hydrogels will be used as biomaterial scaffold systems mimicking the extracellular matrix of native tissues. This process consists of preparing mESCs in a polymerizable PEG diacrylate (PEGDA) solution at different final concentrations such as 15% and 20%. Our goal is that these efforts will lead to three applications: (a) understand the differentiation of mouse embryonic stem cells (mESCs) in microenvironments, ideal for the building of cellular machines (b) employ biology and engineering principles to determine the differentiation of mESCs into different cell types, and (c) determine the viability of stem cells under three-dimensional encapsulation of cells.