Generation of Precision Cut-Lung slices (PCLS) for ex vivo modeling of lung disease

Procedure

Lung tissue of animal or human origin (healthy or diseased) is filled with agarose (1.5-3 w/v %) and allowed to solidify at 4 °C. Regions of interest are then dissected and sliced using a vibrating microtome (7000SMZ-2 Vibrotome, Campden Instruments Ltd.) at thickness of 300-500 µm. The PCLS can then be maintained ex vivo for 5-7 days without major losses in cell viability and native tissue characteristics. During this time window, disease characteristics can be investigated (inherent or induced) and pharmacological approaches evaluated. One of the many advantages of using PCLS for pulmonary research is the possibility to perform microscopy-based imaging with high spatiotemporal resolution (see figure below).

Specific Examples

Our lab utilize ex vivo culture of PCLS for several purposes. One example is disease modeling of lung fibrosis; this is accomplished by treatment of PCLS with a fibrosis cocktail which induces an early stage-like fibrotic response within 48 hours. This model can be used to study both mechanisms involved in disease progression and pharmacological interventions. Recently, our lab has additionally started using PCLS-based models to study acute lung diseases, such as infection by bacteria (in collaboration with Drs. Flávia Viana and Gunnar Schroeder at Queen’s University Belfast, Northern Ireland) or viruses (in collaboration with Dr. Lena Uller at Lund University, Sweden).