PDMS microstencil plate-supported fabrication of ultra-thin, condensed ECM membranes for separated cell coculture on both surfaces

Thin membrane-based cell culture platforms have recently gained much attention for facilitating separated but adjacent cocultures of heterotypic cells in in vivo tissue-mimetic microenvironments. Here we propose a new approach to preparing significantly thin but highly stable membranes composed of extracellular matrix (ECM) components, mainly type I collagen, that are supported by PDMS microstencil plates. A collagen solution was introduced into the through holes of a microstencil plate and dried to form condensed membranes with a thickness of less than 1 mu m. A coculture of HepG2 and Swiss-3T3 cells was performed on both membrane surfaces, and gene expression assays revealed the superiority of the presented thin membranes. Release of the membranes from the plate was possible, as was the integration of the membranes into microfluidic systems to perform perfusion cultures of cells. We were also able to prepare Matrigel membranes by the same procedure. The presented thin membrane-based cell culture platforms would provide physiologically suitable conditions for cells and thus would be widely applicable to a variety of in vitro cell culture systems.