PMID- 34096923
OWN - NLM
STAT- MEDLINE
DCOM- 20211004
LR  - 20211117
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 171
DP  - 2021 May 24
TI  - Precision Cut Lung Slices as an Efficient Tool for Ex vivo Pulmonary Vessel 
      Structure and Contractility Studies.
LID - 10.3791/62392 [doi]
AB  - The visualization of murine lung tissue provides valuable structural and cellular 
      information regarding the underlying airway and vasculature. However, the 
      preservation of pulmonary vessels that truly represents physiological conditions 
      still presents challenges. In addition, the delicate configuration of murine 
      lungs result in technical challenges preparing samples for high-quality images 
      that preserve both cellular composition and architecture. Similarly, cellular 
      contractility assays can be performed to study the potential of cells to respond 
      to vasoconstrictors in vitro, but these assays do not reproduce the complex 
      environment of the intact lung. In contrast to these technical issues, the 
      precision-cut lung slice (PCLS) method can be applied as an efficient alternative 
      to visualize lung tissue in three dimensions without regional bias and serve as a 
      live surrogate contractility model for up to 10 days. Tissue prepared using PCLS 
      has preserved structure and spatial orientation, making it ideal to study disease 
      processes ex vivo. The location of endogenous tdTomato-labeled cells in PCLS 
      harvested from an inducible tdTomato reporter murine model can be successfully 
      visualized by confocal microscopy. After exposure to vasoconstrictors, PCLS 
      demonstrates the preservation of both vessel contractility and lung structure, 
      which can be captured by a time-lapse module. In combination with the other 
      procedures, such as western blot and RNA analysis, PCLS can contribute to the 
      comprehensive understanding of signaling cascades that underlie a wide variety of 
      disorders and lead to a better understanding of the pathophysiology in pulmonary 
      vascular diseases.
FAU - Klouda, Timothy
AU  - Klouda T
AD  - Divisions of Pulmonary Medicine, Boston Children's Hospital.
FAU - Kim, Hyunbum
AU  - Kim H
AD  - Divisions of Pulmonary Medicine, Boston Children's Hospital.
FAU - Kim, Jiwon
AU  - Kim J
AD  - Divisions of Pulmonary Medicine, Boston Children's Hospital.
FAU - Visner, Gary
AU  - Visner G
AD  - Divisions of Pulmonary Medicine, Boston Children's Hospital.
FAU - Yuan, Ke
AU  - Yuan K
AD  - Divisions of Pulmonary Medicine, Boston Children's Hospital; 
      ke.yuan@childrens.harvard.edu.
LA  - eng
GR  - R01 HL150106/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20210524
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - *Lung
MH  - *Lung Diseases
MH  - Mice
MH  - Muscle Contraction
MH  - Signal Transduction
MH  - Specimen Handling
EDAT- 2021/06/08 06:00
MHDA- 2021/10/05 06:00
CRDT- 2021/06/07 12:23
PHST- 2021/06/07 12:23 [entrez]
PHST- 2021/06/08 06:00 [pubmed]
PHST- 2021/10/05 06:00 [medline]
AID - 10.3791/62392 [doi]
PST - epublish
SO  - J Vis Exp. 2021 May 24;(171). doi: 10.3791/62392.