Multilayer nanofilms, formed by the layer-by-layer (LbL) assembly of positively and negatively charged polyelectrolytes, are promising candidates as cell contacting biomaterials. We investigate here the influence of film properties – composition, charge, rigidity, biofunctionality – on the attachment and viability of human hepatocellular carcinoma (HepG2) and human fetal liver (HFL) cells. We employ polymers of biological origin (polysaccharides, polyamino acids) and control film properties (thickness, viscous modulus, elastic modulus) through polymer type, solution salt concentration, and post-formation chemical cross-linking steps. In situ characterization is performed via quartz crystal microgravimetry with dissipation (QCMD) and optical waveguide lightmode spectroscopy (OWLS). We find film composition and rigidity to be the most important variables in promoting growth and function of both HepG2 and HFL cells. In contrast, film charge and biofunctionality appear have a lesser effect. The best performing systems uncovered here are excellent candidates for liver tissue engineering applications.