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[Isolation and observation of ciliated cells from guinea pig's nasal epithelium]
Objective:
To investigate nasal physiological function at a cellular level and to establish a method for isolating single ciliated cells from guinea pig's nasal epithelium.
Methods:
The ciliated cells were isolated by means of mechanical dissociation after papain treatment and a differential interference contrast (DIC) microscope was used for 1000-fold observation (objective lens: 100-fold; eye piece: 10-fold).
Results:
A total of 12,587 single ciliated cells could be isolated from nasal epithelium of each guinea pig. The criteria for good viability of isolated nasal ciliated cells include: (1) actively motile cilia; (2) a smooth cytoplasmic membrane; (3) cytoplasm in a state of translucence (birefringence); (4) cells neither swollen nor shrunk. With short term culture at room temperature, the isolated nasal ciliated cells retained a good viability for 2-9 hours or longer. The living ciliated cells showed swing motions of cilia with fast and slow phases at a rate of 51-119 times per minute.
Conclusion:
A method for isolating ciliated cells from guinea pig's nasal epithelium and a criteria for good viability of isolated ciliated cells have been established. Ciliary motion pattern and ciliary beating frequency were also studied.