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Gerckens, M. ; Alsafadi, H.N.* ; Wagner, D.E.* ; Lindner, M.* ; Burgstaller, G. ; Königshoff, M.

Generation of human 3D lung tissue cultures (3D-LTCs) for disease modeling.

J. Vis. Exp.:e58437 (2019)
Publ. Version/Full Text DOI
Open Access Gold (Paid Option)
Creative Commons Lizenzvertrag
Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) used as 3D lung tissue cultures (3D-LTCs) represent an elegant and biologically highly relevant 3D cell culture model, which highly resemble in situ tissue due to their complexity, biomechanics and molecular composition. Tissue slicing is widely applied in various animal models. 3D-LTCs derived from human PCLS can be used to analyze responses to novel drugs, which might further help to better understand the mechanisms and functional effects of drugs in human tissue. The preparation of PCLS from surgically resected lung tissue samples of patients, who experienced lung lobectomy, increases the accessibility of diseased and peritumoral tissue. Here, we describe a detailed protocol for the generation of human PCLS from surgically resected soft-elastic patient lung tissue. Agarose was introduced into the bronchoalveolar space of the resectates, thus preserving lung structure and increasing the tissue's stiffness, which is crucial for subsequent slicing. 500 mu m thick slices were prepared from the tissue block with a vibratome. Biopsy punches taken from PCLS ensure comparable tissue sample sizes and further increase the amount of tissue samples. The generated lung tissue cultures can be applied in a variety of studies in human lung biology, including the pathophysiology and mechanisms of different diseases, such as fibrotic processes at its best at (sub-)cellular levels. The highest benefit of the 3D-LTC ex vivo model is its close representation of the in situ human lung in respect of 3D tissue architecture, cell type diversity and lung anatomy as well as the potential for assessment of tissue from individual patients, which is relevant to further develop novel strategies for precision medicine.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Medicine ; Issue 144 ; 3d Tissue Culture ; Ex-vivo Tissue Culture ; Precision-cut Lung Slices ; Pcls ; 3d-ltcs ; Lung Disease ; Disease Animal Models ; Human Ex-vivo Models ; Lung Fibrosis; Cut Liver Slices; Animal-models; Precision; Fibrosis; Prevalence; Mortality; Airways; Induce; Repair; Pcls
ISSN (print) / ISBN 1940-087X
e-ISSN 1940-087X
Quellenangaben Volume: , Issue: 144, Pages: , Article Number: e58437 Supplement: ,
Publisher JoVE
Publishing Place 1 Alewife Center, Ste 200, Cambridge, Ma 02140 Usa
Reviewing status Peer reviewed