中国

Publications

Single-cell genomics of post-pneumonectomy peripheral lung regeneration

Bennett, R., Ysasi, A., Belle, J. et al.

Introduction
In most mammals, including humans, unilateral pneumonectomy (PNX) results in non-uniform growth of the remaining lung. To identify regions of active regeneration, we used laser microdissection, microfluidics isolation and single-cell microgenomic PCR. 

Methods
C57BL/6 mice, 811 weeks, underwent left PNX. Lungs were harvested on days 1, 3 & 7 after PNX. Lung sections (300 microns) were laser microdissected & enzymatically dissociated. Cells were isolated on an integrated microfluidics chip (Fluidigm). Custom PCR arrays were used to investigate expression of 96 genes implicated in lung regeneration.

Results
Laser microdissection yielded 13 x 103 cells/alveolar duct. Single-cell analysis revealed gene clusters corresponding to previously identified cell types (e.g., endothelial, myofibroblasts). Comparison of gene expression between cells isolated on the C1 chip vs by flow cytometry showed variable expression in some genes post-PNX that was not evident in population-based flow cytometry analysis. Cell-type specific population dynamics were also notable over the first 7 days post-PNX. 

Conclusion
Laser microdissection facilitates harvesting of regenerating alveolar ducts. Single-cell transcriptional analysis identified individual cell types involved in lung regeneration & highlighted specific cell-type population dynamics within discreet regenerating areas.

Citation

Bennett, R., Ysasi, A., Belle, J. et al. "Single-cell genomics of post-pneumonectomy peripheral lung regeneration" The FASEB Journal (2015): 876.12