Liver cells culture on three-dimensional micropatterned polydimethylsiloxane surfaces

Current in vitro cell culture technologies present some limitations as they can't simulate or mimic in vivo situations. Indeed, in vivo cells function in a three-dimensional (3D) structure where they have a close contact with adjacent cells. In this study, human hepatocarcinoma Hep G2 cells were cultured on 3D micropatterned polydimethylsiloxane (PDMS) substrates. Using soft-lithography techniques, arrays of octagonal macropillars (40 μm width, 50 μm spacing, 100 μm height) in a hexagonal arrangement were fabricated to get a high surface/volume ratio to culture cells at a high density. Those 3D substrates were assembled on a PDMS membrane into a bottomless polycarbonate 12-well plate, thus enabling direct oxygenation of the cells by diffusion of atmospheric air through the polymer. We observed remarkable enhancement of cellular growth due to the high oxygen supply. Cells grew up until forming a quite thick cellular multilayer supported by the pillars. Such 3D substrates are likely to be a new design suitable for inducing organ-specific 3D organization in a conventional microplate-based culture format.