Melo-Narváez, C. ; Gölitz, F.* ; Jain, E. ; Gote-Schniering, J.* ; Stoleriu, M.-G. ; Bertrams, W.* ; Schmeck, B.* ; Yildirim, A.Ö. ; Rauen, U.* ; Wille, T.* ; Lehmann, M.
Cold storage of human precision-cut lung slices in TiProtec preserves cellular composition and transcriptional responses and enables on-demand mechanistic studies.
Respir. Res. 26:57 (2025)
BACKGROUND: Human precision-cut lung slices (hPCLS) are a unique platform for functional, mechanistic, and drug discovery studies in the field of respiratory research. However, tissue availability, generation, and cultivation time represent important challenges for their usage. Therefore, the present study evaluated the efficacy of a specifically designed tissue preservation solution, TiProtec, complete or in absence (-) of iron chelators, for long-term cold storage of hPCLS. METHODS: hPCLS were generated from peritumor control tissues and stored in DMEM/F-12, TiProtec, or TiProtec (-) for up to 28 days. Viability, metabolic activity, and tissue structure were determined. Moreover, bulk-RNA sequencing was used to study transcriptional changes, regulated signaling pathways, and cellular composition after cold storage. Induction of cold storage-associated senescence was determined by transcriptomics and immunofluorescence (IF). Finally, cold-stored hPCLS were exposed to a fibrotic cocktail and early fibrotic changes were assessed by RT-qPCR and IF. RESULTS: Here, we found that TiProtec preserves the viability, metabolic activity, transcriptional profile, as well as cellular composition of hPCLS for up to 14 days. Cold storage did not significantly induce cellular senescence in hPCLS. Moreover, TiProtec downregulated pathways associated with cell death, inflammation, and hypoxia while activating pathways protective against oxidative stress. Cold-stored hPCLS remained responsive to fibrotic stimuli and upregulated extracellular matrix-related genes such as fibronectin and collagen 1 as well as alpha-smooth muscle actin, a marker for myofibroblasts. CONCLUSIONS: Optimized long-term cold storage of hPCLS preserves their viability, metabolic activity, transcriptional profile, and cellular composition for up to 14 days, specifically in TiProtec. Finally, our study demonstrated that cold-stored hPCLS can be used for on-demand mechanistic studies relevant for respiratory research.
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Publication type
Article: Journal article
Document type
Scientific Article
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Keywords
3r ; Fibrosis ; Human Lung Models ; Human Precision-cut Lung Slices (hpcls) ; Long-term Cold Storage ; Tiprotec ; Tissue Preservation; Liver Endothelial-cells; Induced Apoptosis; Cultured-hepatocytes; Immune-responses; Hypoxic Injury; Iron Chelators; Test System; Glycine; Protection; Tissue
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Language
english
Publication Year
2025
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0
HGF-reported in Year
2025
ISSN (print) / ISBN
1465-9921
e-ISSN
1465-993X
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Volume: 26,
Issue: 1,
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Article Number: 57
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BioMed Central
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Campus, 4 Crinan St, London N1 9xw, England
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Peer reviewed
POF-Topic(s)
30202 - Environmental Health
Research field(s)
Lung Research
PSP Element(s)
G-501600-005
G-503100-001
G-501600-001
G-505000-007
Grants
Von Behring Roentgen Foundation
Federal Institute for Risk assessment (Bundesinstitut fur Risikobewertung, BfR)
Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)
German Center for Lung Research (DZL)
German Research Foundation
Projekt DEAL
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Erfassungsdatum
2025-04-11