Human lung tissue digestion

To determine the impact of different enzymatic digestion protocols on the cellular yield and phenotype from peripheral human lung tissue.
Conclusion:
Protocols high in dispase cleaved immune markers CD4, CD8, CD69, and CD103 and contributed to an increased monocyte to macrophage yield. Similarly, dispase led to a differential epithelial cell yield, with increased TSPN8+ and ITGA6+ epithelial cells and reduced CD66e+ cells. When compared to collagenase D, collagenase P protocols yielded increased AT1 and AT2 cells and decreased endothelial cells. These results provide a framework for selecting an enzymatic digestion protocol best suited to the scientific question and allow for comparison of studies using different protocols.
Notes:
Protocol 1: Mechanical digestion alone (gentleMACS) Protocol 2: Liberase + Dnase 1 (Disp lo) Protocol 3: Disp + Col D + Dnase 1 (Disp hi) Protocol 4: Col P + Dnase 1 (Col_P) Protocol 5: Col D + Dnase 1 + trypsin in (Col_D) Protocol 6: Col_D + density centrifugation