A thermoresponsive, micro-roughened cell culture surface

Abstract

Surface topography has been shown to play a major role in cell behavior, but has yet to be seriously exploited in the field of cell surface engineering. In the present work, surface roughness has been used in combination with the thermoresponsive polymer polyisopropylacrylamide (PIPAAm) to generate cell sheets with tailored biochemical properties. Micro-roughened polystyrene (PS) with 1.5-5.5 μm features was derivatized with PIPAAm to form a cell culture surface for the growth of human fibroblast cell sheets that exhibit a modified cytoskeleton and extracellular matrix. Fibroblasts cell sheets cultured on the rough surfaces had fewer actin stress fibers and twice the average fibronectin (FN) fibril formation when compared to cell sheets on flat substrates. The cell sheets harvested from the roughened PS were collected after only 2 days of culture and detached from the PIPAAm grafted surface in <1h after cooling the culture system. The simple and rapid method for generating cell sheets with increased FN fibril formation has applications in tissue grafts or wound repair and has demonstrated that the thermoresponsive surface can be used for reliable cell sheet formation.

Keywords: Cell sheet engineering; Cytoskeleton; Extracellular matrix (ECM); Polyisopropylacrylamide (PIPAAm); Surface roughness.