It may be hypothesized that wound healing will benefit from polymer membranes coated with extracellular matrix macromolecules. Here we describe the behavior of dermal fibroblasts on polysulfone (PSU) membranes, and PSU membranes covered with chitosan (Ch), chondroitin sulfate (CS), or hyaluronan, using an additional intermediary ionic charge modification with poly-(acrylonitrile-co-methallyl sulfonate) (AN69), which allows binding of the polysaccharidic macromolecules. Cell adhesion, proliferation, cell signaling, and collagen gene expression were investigated. Ch and CS were found unable to support cell adhesion and proliferation. In contrast, both PSU and hyaluronic acid-coated PSU membranes appeared as suitable materials to culture fibroblasts and support their matrix synthesis capacity. Moreover, they induce type III collagen expression in addition to type I, suggesting that they promote a fetal-like environment that could be beneficial for wound healing.