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In response to an applied pressure, significantly lower aspiration length is observed for untreated contractile cells compared to cells in which actin polymerisation is chemically inhibited, demonstrating the important contribution of stress fibres in the biomechanical behaviour of spread cells.
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In addition, spreading appeared to be an all-or-none process, and the fraction of spreading cells was tightly correlated to the spreading rate and spreading area.
Percentage of spreading cells was defined as (spreading cells/total number of adherent) X 100.
Finally, the possibility of combining this cell patterning technique with high definition imaging technologies opens the door to a new generation of single cell studies: the behaviour of single cells (spreading, polarisation, differentiation, division) may be controlled using our micro-patterns and probed using a combination of live imaging and FRET.
The spreading of cells on the resulting patterns and the induction of polarisation using asymmetric islands demonstrate the potential of this technique for controlling the behaviour of single cells.
The characterisation of stress fibres and nucleus biomechanics in spread cells presented in the current study can potentially be used to guide tissue engineering strategies to control cell behaviour and gene expression.
First, after depolymerization of microtubules, spreading cells exhibited morphological oscillations.
The induction of such cytoskeletal polarisation enables cells to maximise their spreading, whilst maintaining a low membrane curvature, in very good agreement with the literature describing the behaviour of other cell types on ECM micro-patterns [42].
The behaviour of each cell is dependent only on the state of its eight immediate neighbours, according to the following rules: Live cells: a live cell with zero or one live neighbours will die.
Instead, we analysed the spatial behaviour of transient, spreading changes in perfusion in the peri-infarct region.
Examples of spread and un-spread cells are shown in Supplementary Figure S1B.
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