On-chip non-contact mechanical cell stimulation - quantification of SKOV-3 alignment to suspended microstructures

Abstract

Although the accumulation of random genetic mutations have been traditionally viewed as the main cause of cancer progression, altered mechanobiological profiles of the cells and microenvironment also play a major role as a mutation-independent element. To probe the latter, we have previously reported a microfluidic cell-culture platform with an integrated flexible actuator and its application for sequential cyclic compression of cancer cells. The platform is composed of a control microchannel in a top layer for introducing external pressure, and a polydimethylsiloxane (PDMS) membrane from which a monolithically-integrated actuator protrudes downwards into a cell-culture microchannel. When actively actuated, the integrated actuator, referred to as micro-piston, transfers the pressure from the control channel as a mechanical force to the cells underneath. When not actuated, the micropiston remains suspended above cells, separated from the latter via a liquid-filled gap of ∼108 μm. Despite the lack of direct physical contact between the micro-piston and cells in the latter arrangement, we observed distinct alignment of SKOV-3 ovarian cancer cells to the piston shape. To charaterize this observation, micro-piston localization, shape, and size were adjusted and the directionality of a mono-layer of SKOV-3 cells relative to the suspended structure probed. Cell alignment analysis was performed in a novel, label-free approach by measuring elongation angles of whole cell bodies with respect to micro-piston peripheries. Alignment of SKOV-3 cells to the structure outline was significant for circular, triangular and square micro-piston when compared to control areas without micro-piston on the same chip. The effect was present irrespective of whether cells were loaded with micro-pistons in static position (∼108 μm gap) or actively retracted using vacuum (>108 μm gap). Similar alignment was not observed for MCF7 cancer cells and MCF10A non-cancerous epithelial cells. The reported observation of directional movement and growth of SKOV-3 cells towards the region under micro-pistons point towards a to-date unexplored mechanotactic behaviour of these cells, warranting future investigations regarding the mechanisms involved and the role these may play in cancer.