Authors: Tamer Badawy, Yukari Kyumoto-Nakamura, Norihisa Uehara, Jingqi Zhang, Hidenobu Hiura, Soichiro Sonoda, Takayoshi Yamaza, Akiko Kukita, Toshio Kukita.
Aim: To obtain suitable immunological tool to identify the terminal differentiation of odontoblasts and for enrichment and purification of functional odontoblasts useful for dentin regeneration.
Methods: A panel of monoclonal antibodies (MAbs) recognizing cell-surface of rat osteoblastic cell line was prepared by utilizing the B cell hybridoma technology. Among MAbs recognizing osteoblasts specifically, odontoblast-reactive MAb was selected for immunofluorescence staining of cryosections of rat mandibles followed by observation by use of confocal laser scanning microscopy.
Results: Molecular size of the antigen recognized by A7 MAb (A7 antigen) was revealed to be around 45 KDa by western blotting analysis. A7 antigen was expressed on cell-surface of unfixed osteoblasts in culture. Immunofluorescence staining and confocal laser analysis demonstrated that A7 antigen is specifically expressed in mature odontoblasts in a typical membrane expression pattern.
Conclusions: A7 antigen was revealed to be a unique cell-surface antigen expressed on osteoblasts and odontoblasts. This novel MAb-A7 should be a quite useful immunological tool for enrichment and purification of functional odontoblasts directly from dental pulp tissues in regenerative dental medicine.
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