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This research aimed to combine 3 cell and tissue culture technologies to obtain mechanistic insights of cells in porous scaffolds. When cultivated on 2D (2-dimensional) surfaces, HDFs (human dermal fibroblasts) behaved individually and had no strict requirement on seeding density for proliferation; while HaCat cells relied heavily on initial densities for proliferation and colony formation, which was facilitated when co-cultured with HDFs. Experiments using a 3D CCIS (3-dimensional cell culture and imaging system) indicated that HDFs colonised open pores of varying sizes (125-420 μm) on modular substrates via bridge structures; while HaCat cells formed aperture structures and only colonised small pores (125 μm). When co-cultured, HDFs not only facilitated HaCat attachment on the substrates, but also coordinated with HaCat cells to colonise open pores of varying sizes via bridge and aperture structures. Based on these observations, a 2-stage strategy for the culture of HDFs and HaCat cells on porous scaffolds was proposed and applied successfully on a cellulosic scaffold. This research demonstrated that cell colonisation in scaffolds was dependent on multiple factors; while the integrated 2D&3D culture technologies and the 3D CCIS was an effective and efficient approach to obtain mechanistic insights of their influences on tissue regeneration.

Mechanistic Insights of Cells in Porous Scaffolds via Integrated Culture Technologies