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Isolation, culture and identification of human odontoclasts

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Author:
No author available
Journal Title:
Chinese Journal of Stomatology
Issue:
6
DOI:
10.3760/cma.j.issn.1002-0098.2013.06.004
Key Word:
牙,乳;细胞培养技术;人破牙细胞;酶消化法;Tooth,deciduous;Cell culture techniques;Human odontoclasts;Enzyme digestion

Abstract: 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.

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