Organoids are three dimensional cellular structures formed by epithelial cells cultured either alone or together with their support cells, the fibroblasts. When placed in the right conditions, epithelial stem cells can proliferate and give rise to differentiated daughter cells which resemble their counterparts in the lung environment. This culture system can be used to expand a cell population to be used for organ regeneration, to better understand how these cells grow and differentiate, and to test therapeutic compounds for lung diseases.
The first step is to break the human or mouse lung tissue into single cells. Then the epithelial cells are separated from other cell types and isolated by flow cytometry. A number of these cells are mixed with fibroblasts (in our assay) and cultured inside a gel-like structure called Matrigel on top of a well insert. Cell culture medium is added to the bottom compartment. Defining the composition of the cell culture medium is a great challange, as different cell types require different conditions to grow.
Roxana Wasnick MD PhD
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