Embryonic stem cells have the potential to differentiate into multiple cell types including insulin-producing cells (IPCs), which is becoming one of the promising cell sources for treating type 1 diabetes mellitus. However, in order to achieve functional stem cell-derived cells, it is important to generate more mature IPCs and to keep long-term viability post differentiation process. In this study, we varied several factors including different embryonic body culture conditions, digested cells seeding density and various coatings required for differentiation to optimize a previously established protocol to enhance the overall differentiation efficiency. Moreover, a three-dimensional in vitro collagen tissue culture system was prepared to provide a more physiological culture environment for stem cell-derived IPCs. Survivability of IPCs was examined under both static and flow conditions and low flow rate of 0.02 ml/min resulted in better survival of IPCs in in vitro three-dimensional tissues
