PMID- 30829341
OWN - NLM
STAT- MEDLINE
DCOM- 20200203
LR  - 20200203
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 144
DP  - 2019 Feb 12
TI  - Generation of Human 3D Lung Tissue Cultures (3D-LTCs) for Disease Modeling.
LID - 10.3791/58437 [doi]
AB  - 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 
      microm 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.
FAU - Gerckens, Michael
AU  - Gerckens M
AD  - Comprehensive Pneumology Center, Ludwig-Maximilians-Universitat and Helmholtz 
      Zentrum Munich; German Center of Lung Research (DZL); Translational Lung Research 
      and CPC-M bioArchive, Helmholtz Zentrum Munchen, Comprehensive Pneumology Center 
      Munich DZL/CPC-M.
FAU - Alsafadi, Hani N
AU  - Alsafadi HN
AD  - Department of Experimental Medical Science, Lung Bioengineering and Regeneration, 
      Lund University; Wallenberg Center for Molecular Medicine, Lund University; Stem 
      Cell Centre, Lund University.
FAU - Wagner, Darcy E
AU  - Wagner DE
AD  - Department of Experimental Medical Science, Lung Bioengineering and Regeneration, 
      Lund University; Wallenberg Center for Molecular Medicine, Lund University; Stem 
      Cell Centre, Lund University.
FAU - Lindner, Michael
AU  - Lindner M
AD  - German Center of Lung Research (DZL); Asklepios Fachkliniken Munich-Gauting.
FAU - Burgstaller, Gerald
AU  - Burgstaller G
AD  - Comprehensive Pneumology Center, Ludwig-Maximilians-Universitat and Helmholtz 
      Zentrum Munich; German Center of Lung Research (DZL); Translational Lung Research 
      and CPC-M bioArchive, Helmholtz Zentrum Munchen, Comprehensive Pneumology Center 
      Munich DZL/CPC-M; gerald.burgstaller@helmholtz-muenchen.de.
FAU - Konigshoff, Melanie
AU  - Konigshoff M
AD  - Comprehensive Pneumology Center, Ludwig-Maximilians-Universitat and Helmholtz 
      Zentrum Munich; German Center of Lung Research (DZL); Translational Lung Research 
      and CPC-M bioArchive, Helmholtz Zentrum Munchen, Comprehensive Pneumology Center 
      Munich DZL/CPC-M; Department of Medicine, Division of Pulmonary Sciences and 
      Critical Care Medicine, University of Colorado; melanie.koenigshoff@ucdenver.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20190212
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Cell Culture Techniques
MH  - Humans
MH  - Lung/*pathology
MH  - Molecular Conformation
EDAT- 2019/03/05 06:00
MHDA- 2020/02/06 06:00
CRDT- 2019/03/05 06:00
PHST- 2019/03/05 06:00 [entrez]
PHST- 2019/03/05 06:00 [pubmed]
PHST- 2020/02/06 06:00 [medline]
AID - 10.3791/58437 [doi]
PST - epublish
SO  - J Vis Exp. 2019 Feb 12;(144). doi: 10.3791/58437.