title: A mineralizing rat dental pulp cell subline expressing collagen type I and dentin sialoprotein-phosphophoryn transcripts.
abstract: 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.
authors.affiliation: Department of Cariology, Restorative Sciences and Endodontics, School of Dentistry, University of Michigan, Ann Arbor 48109-1078, USA. helenar@umich.edu
authors.forename: Helena H
authors.initials: HH
authors.lastname: Ritchie
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Kasugai
authors.forename: Peter
authors.initials: P
authors.lastname: Moller
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism*; D017931:DNA Primers; D003782:Dental Pulp / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D010750:Phosphoproteins / Q000235:genetics*; D011498:Protein Precursors; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D012795:Sialoglycoproteins / Q000235:genetics*; D014074:Tooth Calcification; D000099311:Dentin Sialophosphoprotein
chemical_list: D024042:Collagen Type I; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D011498:Protein Precursors; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D000099311:Dentin Sialophosphoprotein; C017630:phosphophoryn