PMID- 35938685
OWN - NLM
STAT- MEDLINE
DCOM- 20220809
LR  - 20221015
IS  - 2691-1299 (Electronic)
IS  - 2691-1299 (Linking)
VI  - 2
IP  - 8
DP  - 2022 Aug
TI  - The Use of Precision-Cut Lung Slices for Studying Innate Immunity to Viral 
      Infections.
PG  - e505
LID - 10.1002/cpz1.505 [doi]
LID - e505
AB  - Precision-cut lung slices (PCLS) are a novel tool to study cells of the lower 
      airways. As PCLS retain the integrity and architecture of the lung, they 
      constitute a robust model for studying the cells of the lower respiratory tract. 
      Use of PCLS for imaging has been previously documented; however, other 
      applications and techniques can also be applied to PCLS to increase their use and 
      therefore decrease the number of animals needed for each experiment. We present a 
      detailed protocol for generating PCLS from the murine lung. We show that cultured 
      PCLS remain viable up to at least 8 days of culture, that RNA can be isolated 
      from the tissue, and that flow cytometry can be carried out on the cells obtained 
      from the PCLS. Furthermore, we demonstrate that cytokines and chemokines can be 
      detected in the culture supernatants of PCLS exposed to viruses. Overall, these 
      protocols expand the use of PCLS, especially for infection studies. (c) 2022 The 
      Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: 
      Precision-cut lung slices (PCLS) Basic Protocol 2: PCLS culture and viability 
      Basic Protocol 3: RNA isolation from PCLS, cDNA conversion, and RT-qPCR Basic 
      Protocol 4: Staining of cells from PCLS for flow cytometry Basic Protocol 5: In 
      vivo RSV administration and ex vivo PCLS RSV exposure.
CI  - (c) 2022 The Authors. Current Protocols published by Wiley Periodicals LLC.
FAU - Michalaki, Christina
AU  - Michalaki C
AD  - Section of Respiratory Infections, National Heart and Lung Institute, Imperial 
      College London, London, United Kingdom.
FAU - Dean, Charlotte
AU  - Dean C
AD  - Cardio Respiratory Interface Section, National Heart and Lung Institute, Imperial 
      College London, London, United Kingdom.
FAU - Johansson, Cecilia
AU  - Johansson C
AD  - Section of Respiratory Infections, National Heart and Lung Institute, Imperial 
      College London, London, United Kingdom.
LA  - eng
GR  - MR/V000659/1/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PL  - United States
TA  - Curr Protoc
JT  - Current protocols
JID - 101773894
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Animals
MH  - Immunity, Innate
MH  - *Lung
MH  - Mice
MH  - RNA
MH  - Thorax
MH  - *Virus Diseases
PMC - PMC9545600
OTO - NOTNLM
OT  - cytokines
OT  - ex vivo model
OT  - innate immunity
OT  - lower respiratory tract
OT  - lung
OT  - viral infections
COIS- No conflict of interest has been identified.
EDAT- 2022/08/09 06:00
MHDA- 2022/08/10 06:00
PMCR- 2022/10/07
CRDT- 2022/08/08 07:23
PHST- 2022/08/08 07:23 [entrez]
PHST- 2022/08/09 06:00 [pubmed]
PHST- 2022/08/10 06:00 [medline]
PHST- 2022/10/07 00:00 [pmc-release]
AID - CPZ1505 [pii]
AID - 10.1002/cpz1.505 [doi]
PST - ppublish
SO  - Curr Protoc. 2022 Aug;2(8):e505. doi: 10.1002/cpz1.505.