Advanced pulmonary models
The present invention relates to microfluidic fluidic systems and methods for the in vitro modeling diseases of the lung and small airway. In one embodiment, the invention relates to a system for testing responses of a microfluidic Small Airway-on-Chip infected with one or more infectious agents (e.g. respiratory viruses) as a model of respiratory disease exacerbation (e.g. asthma exacerbation). In one embodiment, this disease model on a microfluidic chip allows for a) the testing of anti-inflammatory and/or anti-viral compounds introduced into the system, as well as b) the monitoring of the participation, recruitment and/or movement of immune cells, including the transmigration of cells. In particular, this system provides, in one embodiment, an in-vitro platform for modeling severe asthma as “Severe Asthma-on-Chip.” In some embodiments, this invention provides a model of viral-induced asthma in humans for use in identifying potentially effective treatments.
1. A method of culturing, comprising:
a) providing i) a microfluidic device comprising first and second channels, said first channel comprising lung cells at an air-liquid interface, and ii) a solution of bacteria;
b) introducing said solution of bacteria into said first channel such that said bacteria contact and adhere to said lung epithelial cells to produce adhered bacteria; and
c) removing said solution of bacteria; and
d) culturing said lung epithelial cells and said adhered bacteria at an air-liquid interface in the absence of fluid flow in said first channel to produce viable cultured epithelial cells.
2. The method of claim 1 , wherein said lung epithelial cells are small airway cells.
3. The method of claim 1 , further comprising, prior to step b) removing mucus from said first channel.
4. The method of claim 1 , wherein said solution of bacteria comprises 2-3×10 6 colony forming units (CFUs) in Hank's balanced salt solution (HBSS).
5. The method of claim 1 , wherein said solution of bacteria is in said first channel for 2 hours.
6. The method of claim 1 , further comprising, prior to step d), removing any nonadherent bacteria.
7. The method of claim 1 , further comprising flowing culture media in said second channel, and not in said first channel.
8. The method of claim 1 , wherein said lung epithelial cells are fully differentiated mucociliary bronchiolar airway cells.
9. The method of claim 1 , wherein said bacteria are S. pneumoniae.