A 3D printed device for in vitro generation of stratified epithelia at the air-liquid interface

Benjamin John Hewitt, Joanna Batt, Richard Shelton, Paul Cooper, Gabriel Landini, Robert Lucas, Malgorzata Wiench*, Michael Milward*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Air-liquid interface (ALI) cultures are used to produce stratified epithelial tissues in vitro, notably for the production of oral mucosal equivalents. Currently, there are few purpose-built devices which aim to enhance the ease and reproducibility of generating such tissue. Most ALI cultures utilise stainless steel grids or cell culture inserts to elevate the matrix or scaffold to the surface of the culture media. Here, a novel buoyant epithelial culture device (BECD) was designed to both contain a fibroblast-seeded collagen hydrogel and float in culture media, thereby automatically maintaining the ALI without further user intervention. BECDs aim to mitigate several issues associated with ALI culture; reducing the chance of media flooding the epithelial layer from physical disturbance, reducing technique-sensitivity for less experienced users, and improving the reproducibility of the epithelia generated. H400 oral squamous cell carcinoma cells cultured in BECDs for 7, 14 and 21 days showed continuous increase in epithelial tissue thickness with expected localisation of epithelial differentiation markers: cytokeratin 5, involucrin and E-cadherin. Fused filament fabrication 3D printing with polypropylene used in BECD production allows for rapid turnover and design iteration, presenting a versatile, adaptable and useful tool for application in in vitro cell culture.
Original languageEnglish
Pages (from-to)599-609
Number of pages11
JournalTissue Engineering Part C Methods
Issue number11
Early online date1 Sept 2022
Publication statusPublished - 10 Nov 2022

Bibliographical note

Funding Information
This work was jointly supported by the Biotechnology and Biological Sciences Research Council (BBSRC; reference BB/T508329/1), and GlaxoSmithKline as part of an iCASE PhD studentship.


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