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LONG TERM CULTURE OF PSEUDOSTRATIFIED TRACHEAL BRONCHIAL EPIAIRWAY™ TISSUES.
Hayden, P., Jackson, G.R.
MatTek Corporation, Ashland, MA, United States.
Internal results, (2006).
Keywords: AIR-100-ASY-MM, EpiAirway, Histology, Long term culture, MTT viability assay kit (MTT-100), Potential Difference (PD), Transepitheilial electrical resistance (TER)
Endpoints: Histology, MTT Viability assay, Transepitheilial electrical resistance (TER)
Materials Tested: EpiAirway
Summary:
The highly differentiated EpiAirway tissue model is used to perform inhaled drug delivery, inhalation toxicity, and airway inflammation/irritancy studies, amongst others.
For many applications, it is desirable to maintain the organotypic EpiAirway cultures for extended periods. However, for these studies to be useful, it is important to know whether the properties of the EpiAirway tissues significantly change during extended culture periods.
For this reason, we investigated the EpiAirway properties over a 27 day span.
EpiAirway tissues (AIR-100) taken from standard production batches were packaged, stored overnight at 4ºC (to simulate shipping conditions), and re-equilibrated at 37ºC for 24 hours. The tissues were fed every other day with 5.0 ml of EpiAirway maintenance medium (AIR-100-MM) and the apical surface of the tissue was gently rinsed once each week using PBS. Every other day, the transelectrical resistance (TER) was measured, samples were taken for histology, and the MTT tissue viability was determined.
Throughout the 27 day culture period, TER values remained above 200 Ohm*cm2, a pseudostratified epithelium with columnar cells and well-developed cilia was maintained, and the MTT assay showed no sign of decreased viability.
These results indicate that the EpiAirway tissue will likely prove useful in chronic, long term experiments.
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