Chemically detachable polyelectrolyte multilayer platform for cell sheet engineering

Human gingival fibroblasts (HGFs) cell sheets have a potential use for in vivo wound healing due to the ability of HGFs to adopt a contractile phenotype which is typically expressed during extracellular matrix tissue remodeling. For this purpose, we developed a chemically detachable platform based on poly(allylamine hydrochloride)/poly(styrene sulfonate) multilayer film built on a sacrificial precursor film which served as a substrate for HGF cell layer formation. The sacrificial precursor film, based on disulfide-containing polycation and polyanion, is degradable under mild conditions compatible for cell sheet detachment. Cellular viability and cell phenotype analysis of HGF show that the designed platform promotes cell phenotype switch into contractile phenotype, maintained after cell sheet lift-off. This contractile phenotype is acquired by fibroblasts during in vivo wound healing and tissue remodeling. HGFs cell sheet fragments, obtained by this detachment process, could be cultured later on showing a good retention of the typical spindle-shape of differentiated cells after 10 days of culture. HGFs cell sheets have great potential applications as autologous substrates for tissue repair and cellular synthetic platforms for research on connective tissue diseases or evaluation of novel therapeutic agents.