A mineralizing rat dental pulp cell subline expressing collagen type I and dentin sialoprotein-phosphophoryn transcripts
A mineralizing rat dental pulp cell subclone, termed M2H4, was selected from single-cell cloning of the rat dental pulp cell line RPC-C2A by screening confluent single-cell cultures for their ability to undergo mineralization. To induce mineralization, confluent single-cell cultures were treated for 8 d with ascorbic acid followed by the addition of inorganic phosphate to a final concentration of 4 mM for an additional 3 d. Confluent M2H4 subclones were shown by immunofluorescence and electron microscopy to form collagen type I fibrils. Furthermore, using reverse transcriptase-polymerase chain reaction, this subclone was found to be capable of expressing dentin sialoprotein-phosphophoryn (DSP-PP) transcripts, an odontoblast-specific marker. Thus, this newly identified mineralizing rat M2H4 subclone possesses odontoblast-like characteristics and can serve as an in vitro model for examining the role of DSP and PP in the formation of mineralized dentin.
Key wordsdentin odontoblasts ascorbic acid dentin sialoprotein-phosphophoryn transcript mineralization
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