Fabrication and characterization of egg white cryogel scaffold for three-dimensional (3D) cell culture

Abstract Three-dimensional (3D) cell culture models represent the cell-cell and cell-ECM interactions and offer reliable data than monolayer cell culture systems. In 3D models scaffolds are used to reflect the microenvironment which mimics cells native condition. The present study focused on fabrication of a cost-effective scaffold system for 3D cell culture. The cost-affordable and bio-active egg white (EW) was used as a polymer to make the scaffold by adopting cryogelation process, in which different concentrations (0.25%, 0.5%, 0.75% and 1%) of glutaraldehyde solution was used as the cross linker. Scanning Electron Microscopy (SEM) was adopted to analyze the surface morphology and pore size distribution of cryogel. The FT-IR analysis was carried out to analyze the functional groups formed during the cross linking. The water uptake capacity and porosity were measured to analyze the biophysical features of the cryogel. Epithelial cell lines HepG2 and MCF7 were cultured in this cryogel scaffold in 3D format as well as in the conventional 2D format. Cell viability and proliferation efficiency over defined periods were analyzed by MTT assay. The cultured cells in the scaffold were stained using Hoechst and examined in a microscope. The data suggest that the egg white-derived bio-scaffold can be effectively used in 3D culture in view of its evident advantages i.e., homogenous size of pores, amide linkages during cross linking, and the high degree of porosity. Thus, successful development of hen's EW as a cost-affordable cryogel scaffold for 3D culture of cells for use in toxicology and pharmacology is reported.