Integrated Mucus Transport Activities of Airway Epithelia

The UNC CF Center has extensive experience measuring the integrated pulmonary defense activity of mucus transport in both culture and small animal models.

With respect to culture, we have long had in-house our “classic” system whereby cell cultures grown in 1.2 mm Transwell ALI culture systems develop spontaneous rotational mucus transport that can be quantitated by time-lapse fluorescence videomicroscopy (see Figure 13). These techniques have been supplemented recently with novel techniques designed to utilize in-house built culture systems to measure mucus transport rates in linear systems.

The in vitro systems are complemented by a number of techniques to measure mucus transport in mouse models. These techniques include direct deposition of radiotracers or fluorescence tracers, followed by measurement of linear clearance rates with time in the trachea (see Figure 14) or nasopharynx (Figure 15). In addition, techniques have been developed to deposit aerosols on pulmonary surfaces to measure whole lung mucus clearance, using external technetium radioisotope counting techniques (see Figure 16).


Figure 13

Figure 13. Time lapse fluorescence microscopy of air-liquid interface culture surface to measure mucus transport rates.

Figure 15

Figure 15. Mucociliary clearance in the murine posterior nasopharynx (soft palate) following aspiration of 6um fluorescent beads.

Figure 14

Figure 14. Mucociliary clearance (MCC) in the murine trachea following aspiration of fluorescent beads (6 um) in perflurocarbon (0.25 ul/g). The trachea of the anesthetized mouse is exposed and covered with water equilibrated mineral oil for imaging. The first images of MCC are obtained while the mouse is alive (the breathing motions are obvious) and then the anesthetized mouse is euthanized by exsanguination and MCC is again recorded. Images are obtained by placing the mouse under a fluorescent dissection microscope (Leica) and recording with a DAGE MTI video camera.

Figure 16

Figure 13. Time lapse fluorescence microscopy of air-liquid interface culture surface to measure mucus transport rates.

 

 

 

 

 

 

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Inquiries

Questions? If you are interested in learning more about how the UNC CF Center can help further your research, or if you are interested in collaborating with us, please contact our CF Center Director, Dr. Richard Boucher:

7011 Thurston-Bowles Bldg.
The University of North Carolina at Chapel Hill
Campus Box #7248
Phone: (919) 966-1077
Fax: (919) 966-5178
Email: richard_boucher@med.unc.edu