The culture of keratinocytes on flexible membranes has been proposed as a means to simplify, accelerate and improve the efficiency with which proliferating cells are delivered to full thickness or non-healing skin defects. The purpose of this article was to study the ability of chitosan-gelatin manbranes to facilitate the growth of human keratinocytes. The membranes with different chitosan contents were studied. The surface properties of chitosan-gelatin membranes were investigate by SEM, and water contact angle test. The mechanical property of the membranes was tested. Data implied that gelatin could make the membranes more flexible and hydrophilic than chitosan membranes, which may regulate the seeded cells behavior. Loading human keratinocytes on chitosan-gelatin membranes, cells attachment, spread, and growth were investigated by light microscopy, SEM, and MTT test. The results suggested that the adhesion and proliferation of keratinocytes seeded on chitosan-gelatin membranes were same as on tissue culture plate, in which gelatin could modify the interaction between keratinocytes and chitosan membranes. Therefore, chitosan-gelatin membrane is a good candidate for keratinocytes delivery system. (C) 2003 Kluwer Academic Publishers.