PMID- 35129166
OWN - NLM
STAT- MEDLINE
DCOM- 20220405
LR  - 20230511
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 179
DP  - 2022 Jan 21
TI  - Engineered Lung Tissues Prepared from Decellularized Lung Slices.
LID - 10.3791/63151 [doi]
AB  - There is a need for improved 3-dimensional (3D) lung models that recapitulate the 
      architectural and cellular complexity of the native lung alveolus ex vivo. 
      Recently developed organoid models have facilitated the expansion and study of 
      lung epithelial progenitors in vitro, but these platforms typically rely on mouse 
      tumor-derived matrix and/or serum, and incorporate just one or two cellular 
      lineages. Here, we describe a protocol for generating engineered lung tissues 
      (ELTs) based on the multi-lineage recellularization of decellularized 
      precision-cut lung slices (PCLS). ELTs contain alveolar-like structures 
      comprising alveolar epithelium, mesenchyme, and endothelium, within an 
      extracellular matrix (ECM) substrate closely resembling that of native lung. To 
      generate the tissues, rat lungs are inflated with agarose, sliced into 450 
      microm-thick slices, cut into strips, and decellularized. The resulting acellular ECM 
      scaffolds are then reseeded with primary endothelial cells, fibroblasts, and 
      alveolar epithelial type 2 cells (AEC2s). AEC2s can be maintained in ELT culture 
      for at least 7 days with a serum-free, chemically-defined growth medium. 
      Throughout the tissue preparation and culture process, the slices are clipped 
      into a cassette system that facilitates handling and standardized cell seeding of 
      multiple ELTs in parallel. These ELTs represent an organotypic culture platform 
      that should facilitate investigations of cell-cell and cell-matrix interactions 
      within the alveolus as well as biochemical signals regulating AEC2s and their 
      niche.
FAU - Leiby, Katherine L
AU  - Leiby KL
AD  - Department of Biomedical Engineering, Yale University; Yale School of Medicine.
FAU - Ng, Ronald
AU  - Ng R
AD  - Department of Biomedical Engineering, Yale University.
FAU - Campbell, Stuart G
AU  - Campbell SG
AD  - Department of Biomedical Engineering, Yale University; Department of Cellular and 
      Molecular Physiology, Yale School of Medicine.
FAU - Niklason, Laura E
AU  - Niklason LE
AD  - Department of Biomedical Engineering, Yale University; Department of 
      Anesthesiology, Yale School of Medicine; laura.niklason@yale.edu.
LA  - eng
GR  - F30 HL143880/HL/NHLBI NIH HHS/United States
GR  - T32 GM136651/GM/NIGMS NIH HHS/United States
GR  - U01 HL145567/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 - 20220121
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - *Endothelial Cells
MH  - Extracellular Matrix/chemistry
MH  - Lung
MH  - Mice
MH  - Pulmonary Alveoli
MH  - Rats
MH  - *Tissue Scaffolds/chemistry
PMC - PMC10171469
MID - NIHMS1896742
EDAT- 2022/02/08 06:00
MHDA- 2022/04/06 06:00
PMCR- 2023/05/10
CRDT- 2022/02/07 08:43
PHST- 2022/02/07 08:43 [entrez]
PHST- 2022/02/08 06:00 [pubmed]
PHST- 2022/04/06 06:00 [medline]
PHST- 2023/05/10 00:00 [pmc-release]
AID - 10.3791/63151 [doi]
PST - epublish
SO  - J Vis Exp. 2022 Jan 21;(179):10.3791/63151. doi: 10.3791/63151.