Human alveolar epithelial type II cells in healthy and IPF lung

What is an alveolar epithelial cells? Are there subtypes of AEC2s in the human lung? How are they replenished? And how does everything change in IPF. I started with flow cytometry characterization, based on known markers. Next I performed microarray transcriptomic analysis on sorted human AEC2s from normal and IPF lungs. I found new markers and identified new processes that improved our understanding of the human lung.

Epithelial 3D organoids

To study epithelial cell regenerative capacity in vitro, I set-up a co-culture system comprising of lung epithelial cells and lung fibroblast. This was based on the experience gained during my PhD, when I developed an organoid assay for mouse epithelial cells. Defining and refining the culture conditions was a team effort. After many tries, we understood that organoid culture has three phases, each with its own culturing conditions; initiation, expansion and differentiation. This dual co-culture system was used to study both epithelial and fibroblast cells in normal and diseased lung, as well as their interaction. The system is amenable to medium to high through-put upscaling for compound testing and drug discovery.

Precision cut lung slices

The lung is a very complex mixture of cells, which exist in a tight community. They rely on each other for the production of common goods, which they need for their daily existence. We set up an assay where pieces of donated lung (healthy or diseased) were sliced in thin slices and kept in a cell culture dish. We were interested in a particular  in AEC2s, as they seem to be affected in IPF, so I developed methods of culturing and studying AEC2s in PCLS.

Roxana Wasnick MD PhD 

Medical Scientist

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