Data from ameloblast cell lines cultured in 3D using various gel substrates in the presence of ameloblastin.

This article contains data related to the research article in this issue titled ameloblastin promotes polarization of ameloblast cell lines in a 3D cell culture system (Visakan et al., 2022). In the process of amelogenesis, the organic matrix components are pivotal to the establishment of ameloblast-matrix adhesion. Here we employ immortalized ameloblast cell lines and analyse their morphological changes in 3D cell culture when cultured in the presence of recombinant enamel matrix proteins- ameloblastin and amelogenin compared with controls. The recombinant proteins that were purified using high-performance liquid chromatography (HPLC) were characterized using SDS-gel electrophoresis. A 3D-on-top culture technique was employed, and the cells were analysed 24 and 72 h post inoculation using fluorescent and confocal microscopy for qualitative and quantitative changes. Aspect ratio of cells was measured and used as the parameter to compare between test proteins and controls. Repeated measurements of aspect ratio were recorded to analyse for statistical significance. Additionally, three distinct gel substrates were studied to examine the effect of composition and stiffness of the substrate on cell behaviour. The cells in the 3D culture were fixed and labelled using antibodies to junctional complex, polarity and tight junctional proteins following protocols for whole culture fixation. Co-localization between membrane and specific antibody labels were examined under confocal microscopy.