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Research Article | Open Access

The empowering influence of air-liquid interface culture on skin organoid hair follicle development

Jane Sun1, Imaan Ahmed1, Jason Brown2,3, Kiarash Khosrotehrani1,‡( ), Abbas Shafiee1,2,3,‡ ( )
Frazer Institute, Faculty of Medicine, The University of Queensland, Brisbane, QLD, 4102 Australia
Herston Biofabrication Institute, Metro North Hospital and Health Service, Queensland Health, Brisbane, QLD, 4029 Australia
Royal Brisbane and Women's Hospital, Metro North Hospital and Health Service, Queensland Health, Brisbane, QLD, 4029 Australia

‡Kiarash Khosrotehrani and Abbas Shafiee contributed equally as senior authors.

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Highlights

• Generation of human-induced pluripotent stem cells-derived skin organoids is optimized using an air-liquid interface model and compared with theconventional floating culture in low-attachment plates.

• Air-liquid interface culture advances the development of mature skin organoids with improved morphogenesis compared to floating culture.

• Air-liquid interface culture produces skin organoids with increased hair follicle numbers than floating culture.

• This study offers new insights into optimizing skin organoid models to enhance hair follicle development and dermatological research.

Abstract

Background

Rodent models have been widely used to investigate skin development, but do not account for significant differences in composition compared to human skin. On the other hand, two-dimensional and three-dimensional engineered skin models still lack the complex features of human skin such as appendages and pigmentation. Recently, hair follicle containing skin organoids (SKOs) with a stratified epidermis, and dermis layer have been generated as floating spheres from human-induced pluripotent stem cells (hiPSCs).

Methods

The current study aims to investigate the generation of hiPSCs-derived SKOs using an air-liquid interface (ALI) model on transwell membranes (T-SKOs) and compares their development with conventional floating culture in low-attachment plates (F-SKOs).

Results

Mature SKOs containing an epidermis, dermis, and appendages are created in both T-SKO and F-SKO conditions. It was found that the hair follicles are smaller and shorter in the F-SKO compared with T-SKOs. Additionally, the ALI conditions contribute to enhanced hair follicle numbers than conventional floating culture.

Conclusions

Together, this study demonstrates the significant influence of transwell culture on the morphogenesis of hair follicles within SKOs and highlights the potential for refinement of skin model engineering for advancing dermatology and skin research.

Keywords

Air-liquid interfaceDermal papillaPluripotent stem cellRegenerative medicineSkin appendages

References

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Burns & Trauma
Volume 13 Issue 2,
February 2025
Article number: tkae070
DOI: 10.1093/burnst/tkae070

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Cite this article:
Sun J, Ahmed I, Brown J, et al. The empowering influence of air-liquid interface culture on skin organoid hair follicle development. Burns & Trauma, 2025, 13(2): tkae070. https://doi.org/10.1093/burnst/tkae070

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Received: 19 May 2024
Revised: 25 October 2024
Accepted: 30 October 2024
Published: 10 October 2026
© The Author(s) 2025. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.