Multilayer parylene-C stencils for dynamically controlling cell interactions

Most cell processes involve dynamic interactions between the cells and their microenvironment and the ability to control the microenvironment is of crucial importance for studying these processes in-vitro. In this study, we describe a technology for creating multilayer and mechanically robust parylene-C stencils and demonstrate the ability to generate a series of at least 5 temporally controlled cell co-culture micropatterns using embryonic stem cells as the primary cell type. Utilizing layer by layer coatings of collagen and fibronectin, both untreated and anti-stiction layer treated parylene-C surfaces are rendered adhesive for cells. Co-culturing using parylene-C stencils may find broad applications in studies investigating cellular interactions and in studies requiring a controlled microenvironment, such as stem cell differentiation.