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DOI: 10.3791/3261-v
This method allows characterization of extended bacterial co-culture with EpiAirways, a biologically relevant in vitro model using primary human respiratory epithelial tissue. The approach can be utilized with any microbe suitable for long-term co-culture.
This method allows characterization of extended bacterial co-culture with EpiAirways, primary human respiratory epithelial tissue grown at the air-liquid interface, a biologically relevant in vitro model. The approach can be used with any microbe that is amenable to long-term co-culture.
The overall goal of the following experiment is to perform long-term co-culture of Nontypeable Hemophilus influenza or NTHI with primary human respiratory epithelial tissues using the epi airway model. This is achieved by inoculating the apical surface with NTHI to allow the bacteria to adhere to and enter the tissues as a second step. The inserts are washed every 24 hours with prewarm DPBS to mimic normal mucus flow and to remove mucin waste products and any cellular debris.
Next, the tissues are harvested at the desired time point and the survival of either total cell associated or internalized NTHI is determined. Finally, with this method, strain specific characteristics of NTHI survival within the tissues over time based on viable bacteria counts can be evaluated. Though this method can provide insight into long-term infections with nontypeable hemophilus influenza.
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