培养平台影响人肺类器官标志物表达与形态

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• Open access
• Published: 26 October 2026

• Babette Fletemeyer^2 na1,
• Stefanie Dichtl^1 na1,
• Sophie Ann Erckert¹,
• Doris Wilflingseder^2,3 &
• …
• Wilfried Posch  ORCID: orcid.org/0000-0001-8955-7654¹ 

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Abstract

The cultivation platform strongly shapes the development and phenotype of human lung organoids derived from adult stem cells. We compared three systems - animal-derived (AD), animal-free (AF), and scaffold-free (SF) - under basal-out, apical-out, and proximally differentiated conditions. Our goal was to assess how each platform influences epithelial cell-type-associated marker expression and morphology. AD and AF matrices yielded smaller, more uniform organoids, whereas SF cultures produced markedly larger structures, especially in apical-out and proximally differentiated conditions. Bulk RT-qPCR analysis revealed distinct platform-associated marker-expression profiles: the basal cell marker KRT5 peaked in AD basal-out organoids. Under proximal differentiation, KRT5 expression declined across systems, reaching the lowest levels in AF, and SCGB1A1 became undetectable in all systems. In contrast, SF cultures showed the highest expression of the AT1 cell-associated marker AQP5 (basal-out and apical-out) and of the ciliated cell-associated marker FOXJ1 (proximally differentiated), whereas MUC5AC was only slightly upregulated. Notably, ACE2 and TMPRSS2 - key mediators of SARS‑CoV‑2 entry - were induced by proximal differentiation: ACE2 was strongly upregulated in SF organoids, and TMPRSS2 increased under all proximal conditions, independent of platform. Immunofluorescence corroborated mature epithelial phenotypes, revealing robust ciliation across platforms and particularly advanced cilia maturation in AF cultures. Together, these data indicate that the cultivation platform strongly influences lung organoid growth, morphology, and cell-type-associated marker expression. Our findings provide an evidence-based guide for selecting systems to model human respiratory physiology, infection, and regeneration.

Acknowledgements

We thank our technicians Karolin Thurnes, Simon Paulick and Christina Witting (Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, Innsbruck, Austria), for their valuable help and support regarding this study. We would like to express our gratitude to the Hubrecht Laboratory (Hubrecht Organoid Technology, Utrecht, The Netherlands) for providing healthy human lung organoids as well as culture protocols, which were essential for our research. This research was funded in whole by the Austrian Science Fund (FWF) (10.55776/P34070 and 10.55776/PAT5302524 to W.P.).

Funding

This research was funded in whole by the Austrian Science Fund (FWF) (https://doi.org/10.55776/P34070 and https://doi.org/10.55776/PAT5302524 to W.P.).

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Author notes

1. Babette Fletemeyer and Stefanie Dichtl contributed equally to this work.

Authors and Affiliations

1. Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, Innsbruck, Austria

   Stefanie Dichtl, Sophie Ann Erckert & Wilfried Posch

2. Infectiology and Virology Unit, University of Veterinary Medicine Vienna, Vienna, Austria

   Babette Fletemeyer & Doris Wilflingseder

3. Ignaz Semmelweis Institute, Interuniversity Institute for Infection Research, University of Veterinary Medicine Vienna, Vienna, Austria

   Doris Wilflingseder

Authors

1. Babette Fletemeyer
2. Stefanie Dichtl
3. Sophie Ann Erckert
4. Doris Wilflingseder
5. Wilfried Posch

Corresponding authors

Correspondence to Doris Wilflingseder or Wilfried Posch.

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The authors declare no competing interests.

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Fletemeyer, B., Dichtl, S., Erckert, S.A. et al. Culture platform influences marker expression and morphology of human lung organoids. Sci Rep (2026). https://doi.org/10.1038/s41598-026-77318-7

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• Received: 04 December 2025

• Accepted: 19 October 2026

• Published: 26 October 2026

• DOI: https://doi.org/10.1038/s41598-026-77318-7

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