The present study describes the preparation of extracellular matrix (ECM; from porcine omentum) based chitosan composite films for wound dressing applications. The films were prepared by varying the ECM content, whereas, the amount of chitosan was kept constant. The interactions amongst the components of the films were analyzed by FTIR and XRD studies. The films were thoroughly characterized for surface hydrophilicity, moisture retention capability, water vapor permeability, mechanical and biocompatibility. FTIR study indicated that both chitosan and ECM were present in their native form and did not lose their activity. XRD analysis suggested composition dependent change in the crystallinity of the films. The mechanical properties suggested that the composite films had sufficient properties to be used for wound dressing applications. An increase in the ECM content resulted in better hydrophilicity of the films and hence better the moisture retention capacity and retardant water vapor transmission rate property of the composite films. The films were found to be biocompatible to both blood and adipose tissue derived stem cells. In gist, the prepared films may be explored as wound dressing materials. © 2015, Springer Science+Business Media New York.