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The apicobasal polarisation of epithelial cells within an epithelium is critical for its function as a selective barrier. Microenvironmental parameters, including cell-matrix and cell-cell interactions, contribute to the initiation and orientation of this polarity. However, it is often non-trivial to decipher the differential effects of these parameters in a controlled manner using traditional in vitro platforms. A reductionist platform, consisting of E-cadherin coupled onto laterally mobile supported lipid bilayers, was utilised to mimic E-cadherin presentation in the cell membrane. These functionalised bilayers were generated either on flat 2D surfaces or the interior surfaces of round microwells. This platform enabled the study of E-cadherin adhesion and the initiation of polarisation in a controlled environment, where the dimensionality of the microenvironment, type of protein coating and cell shape could be independently studied. A high proportion of single epithelial cells interacted with and clustered cellular E-cadherin in the presence of E-cadherin functionalised bilayers, which was reduced in the presence of integrin-mediated adhesion. The differential response in E-cadherin clustering correlated with the polarisation of E-cadherin and Na,K-ATPase, a reporter for the induction of basolateral polarity. Neither the three-dimensional presentation of E-cadherin nor the cell shape affected E-cadherin clustering or polarisation in single cells. Thus, the mobile presentation of E-cadherin was sufficient to mimic a cell-cell contact and induce basolateral polarisation in single cells.

Microwell arrays have emerged as robust and versatile alternatives to conventional mammalian cell culture substrates. Using standard microfabrication processes, biomaterials surfaces can be topographically patterned to comprise high-density arrays of micron-sized cavities with desirable geometry. Hundreds to thousands of individual cells or cell colonies with controlled size and shape can be trapped in these cavities by simple gravitational sedimentation. Efficient long-term cell confinement allows for parallel analyses and manipulation of cell fate during in vitro culture. These live-cell arrays have already found applications in cell biology, for example to probe the effect of cell colony size on embryonic stem cell differentiation, to dissect the heterogeneity in single cell proliferation kinetics of neural or hematopoietic stem/progenitor cell populations, or to elucidate the role of cell shape on cell function. Here, we highlight the key applications of these platforms, hopefully inspiring biologists to apply these systems for their own studies.