Abstract
Engineering tissue structures that mimic those found in vivo remains a challenge for modern biology. We demonstrate a new technique for engineering composite structures of cells comprising layers of heterogeneous cell types. An acoustofluidic bioreactor is used to assemble epithelial cells into a sheet-like structure. On transferring these cell sheets to a confluent layer of fibroblasts, the epithelial cells cover the fibroblast surface by collective migration maintaining distinct epithelial and fibroblast cell layers. The collective behaviour of the epithelium is dependent on the formation of cell-cell junctions during levitation and contrasts with the behaviour of mono-dispersed epithelial cells where cell-matrix interactions dominate and hinder formation of discrete cell layers. The multilayered tissue model is shown to form a polarised epithelial barrier and respond to apical challenge. The method is useful for engineering a wide range of layered tissue types and mechanistic studies on collective cell migration.
Highlights
Epithelia line the outer surface of organs forming polarised structures that act as barriers to the external environment
We demonstrate that the behaviour of the epithelial cell sheets is critically dependent on the formation of adherens junctions in the acoustic bioreactor; this modifies epithelial cell behaviour so that they act collectively to form a sheet over a fibroblast layer
Exploratory experiments revealed that application of monodisperse bronchial epithelial cells to fibroblast cultures resulted in formation of islands of epithelial cells surrounded by fibroblasts (Fig. 1a,b)
Summary
Epithelia line the outer surface of organs forming polarised structures that act as barriers to the external environment. In the conducting airways of the lungs, the bronchial epithelial surface protects against noxious gases, pathogens and particulates from the inhaled environment[1] An alternative method for creating direct contact co-culture models is through use of cell sheet engineering, a method developed to allow autologous cell sheets to be generated in vitro and transplanted and engrafted in vivo[14] This technique uses a thermoresponsive polymer such as Poly(N-isopropyl acrylamide) which is hydrophobic at 37 °C supporting cell attachment and growth to form a confluent sheet, but hydrophilic at temperatures below 32 °C enabling cell sheet release[15,16,17]. This would offer the primary advantage of thermoresponsive polymers, without the need for exposure to low temperatures or mechanical damage
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