Cholesteryl ester liquid crystals (CELC) were demonstrated with application in biosensing and microtissue regeneration. The affinity of the cells to this liquid crystal is unclear and required further investigation. This study focused on characterising the physical properties of CELC and interaction of human keratinocytes with CELC. The physical properties of CELC were characterised by a custom built contact angle measurement system and bubble pressure measurement apparatus. Other methods such as pendant drop were applied to determine the critical surface tension of the CELC. Then, the characterization of the CELC was continued by using Differential Scanning Calorimeter (DSC), X-ray Diffraction (XRD), Polarising Microscopy (POM) and Fourier Transform Infrared Spectroscopy (FTIR). Nonetheless, the morphology of cells interaction with CELC after it reached confluency was studied using Field Emission Scanning Electron Microscopy (FESEM) and non-contact mode of Atomic Force Microscopy (AFM). The results showed that the critical surface tension of the liquid crystal using contact angle was 37.5 mN/m and the surface tension measured using pendant drop method was found to be 23.6 mN/m. Both results indicate that the surface of the liquid crystal was moderately hydrophobic. From the DSC, CELC was found stable at room and incubator temperature. From XRD results, the compound of CELC interacts in cell culture media self-assembles into lyotropic layer. POM and FTIR analysis showed CELC after immersion in media displayed lyotropic smectic phases. The AFM and FESEM images indicated good adhesion of cells on the CELC. This research thus showed that the hydrophilic layers of lyotropic phase of cholesteryl ester liquid crystal were demonstrated with biophysical properties that support the adhesion of cells