Ion-induced cell sheet detachment from standard cell culture surfaces coated with polyelectrolytes.

Abstract

Polyelectrolyte multilayers (PEMs), formed by alternating layer-by-layer deposition of polyanions and polycations, are an ideal substrate for controlling cellular adhesion and behavior. In the present study we propose a simple mechanism for the controlled detachment of C(2)C(12) myoblasts cell sheets from PEMs consisting of poly(l-lysine) and hyaluronic acid with a topmost layer of fibronectin. The multilayers were deposited on two standard cell culture surfaces: glass and polystyrene. Adding a low concentration of nontoxic ferrocyanide to the cell culture medium resulted in erosion of the polyelectrolyte multilayer and rapid detachment of viable cell sheets. Additional Quartz Crystal Microbalance and Atomic Force Microscopy measurements indicated that the detached cells retained their extracellular matrix and that no polyelectrolyte molecules remained bound to the cell sheets. The dissolution of polyelectrolyte multilayers by multivalent ions is a promising approach to cell sheet engineering that could potentially be used for regenerative medicine.