In this study, the development of a model tracheal mucus with chemical composition and physical properties (bulk viscoelasticity and surface tension) matched to that of native tracheal mucus is described. The mucus mimetics (MMs) were formulated using components that are abundant in tracheal mucus (glycoproteins, proteins, lipids, ions, and water) at concentrations similar to those found natively. Pure solutions were unable to achieve the gel behavior observed with native mucus. The addition of a bifunctional cross-linking agent enabled control over the viscoelastic properties of the MMs by tailoring the concentration of the cross-linking agent and the duration of cross-linking. Three MM formulations with different bulk viscoelastic properties, all within the normal range for nondiseased tracheal mucus, were chosen for investigation of surfactant spreading at the air–mimetic interface. Surfactant spread quickly and completely on the least viscoelastic mimetic surface, enabling the surface tension of the mimetic to be lowered to match native tracheal mucus. However, surfactant spreading on the more viscoelastic mimetics was hindered, suggesting that the bulk properties of the mimetics dictate the range of surface properties that can be achieved. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 1788–1798, 2014.