[Isolation, culture and identification of human odontoclasts].

OBJECTIVE

To isolate, culture and identify odontoclasts in vitro and to establish a method of culturing human odontoclasts.

METHODS

Healthy and retentive deciduous teeth were extracted, and then placed in α-minimum essential medium containing 0.1% collagenase and 0.2% dispase for 1 h.Odontoclasts were obtained and incubated from the absorbing root surfaces of deciduous teeth.Isolated cells were viewed by inverted phase contrast microscope firstly. Then, the isolated odontoclasts were morphologically observed by hematoxylin and eosin staining (HE) and tartrate-resistant acid phosphatase (TRAP) staining. The prepared teeth slices were cocultured with the isolated odontoclasts and scanning electronic microscope(SEM) was used to demonstrate the presence of resorption lacunae.

RESULTS

The isolated odontoclasts appeared as multinucleated giant cell with many vacuolus in cytoplasm. TRAP staining demonstrated that the cytoplasm of the odontoclasts was full of claret-red positive particles.Resorption lacunae on teeth slices which cocultured with odontoclasts were seen under SEM.

CONCLUSIONS

Enzyme digestion is an effective method to isolate odontoclasts from absorbing root surface of deciduous teeth.