Engineering multi-layered tissue constructs using acoustic levitation

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.

Coalescence Dynamics of Acoustically Levitated Droplets

“…Further investigation is required to explore the optimum conditions of droplet coalescence, including the best speed of collision and the combination of liquid properties of the droplets. A clear understanding of droplet dynamics during contactless droplet manipulation via acoustic levitation will provide better physical and application insights for potential lab-on-a-drop applications [28][29][30]. Funding: The authors gratefully acknowledge the support from Japan Society for the Promotion of Science (JSPS) KAKENHI, grant number 15H03925.…”

Coalescence Dynamics of Acoustically Levitated Droplets

“…Further investigation is required to explore the optimum conditions of droplet coalescence, including the best speed of collision and the combination of liquid properties of the droplets. A clear understanding of droplet dynamics during contactless droplet manipulation via acoustic levitation will provide better physical and application insights for potential lab-on-a-drop applications [28][29][30].…”

Assembling Living Building Blocks to Engineer Complex Tissues

“…[202] Notably, ultrasound frequencies have wavelengths that can be used to manipulate microscale entities, such as single cells or cell-laden materials. A common Peer reviewed version of the manuscript published in final form in Advanced Functional Materials (2020) 23/49 example of single cell manipulation is the use of ultrasound standing waves to remotely aggregate cells into sheets, [203] spheroids, [204][205][206] pellet tissue cultures, [207] and organoids. [208] In this regard, acoustic fields can be used either as a technology for modular cellular assembly or as a method for controllably fabricating higher-dimensional living building blocks.…”

“…Acoustic assembly -Single cells [203][204][205][206][207][208][211][212][213][214][215][216][217][218] -Cellularized microcarriers [121] -High throughput and rapid assembly -Can generate complex geometric patterns -No labelling required -Assembly can be impeded by opposing forces (e.g., gravity, acoustic streaming, viscous forces)…”