title: Tissue engineering with dental pulp stem cells:  isolation, characterization, and osteogenic differentiation.
abstract: BACKGROUND
  Dental pulp stem cells (DPSCs) are adult mesenchymal stem cells that have the ability to differentiate into osteoblasts, a fact that is very interesting in the context of tissue engineering. Our purpose was to isolate and characterize DPSCs and to compare the differentiation potential of 3 different osteogenic media.
  
  
  METHODS
  Human dental pulp extracted from healthy young adults was placed in flasks with a mesenchymal expansion medium. At passage 4 DPSCs were analyzed for cell-cycle stage, proliferation, viability, and immunophenotype. DPSCs were grown in 3 different osteogenic media for 40 days. Flasks were incubated at 37 °C in 5% CO2, and the medium was changed twice a week. At day 40, the mineralization of the matrix was determined with Alizarin Red S dye.
  
  
  RESULTS
  After osteogenic induction, DPSCs developed mineralization nodules (clusters), as revealed by Alizarin Red staining. This staining was stronger in the Osteodiff (Miltenyi) medium when compared to the other osteogenic media.
  
  
  CONCLUSIONS
  This study demonstrates the ability of DPSC to differentiate into osteoblasts, especially in the presence of Osteodiff (Miltenyi). DPSCs are therefore a good candidate model for the study of hard-tissue mineralization.
authors.affiliation: Cell Therapy Unit, Hospital Universitario Virgen de la Arrixaca, Faculty of Medicine, University of Murcia, Spain. fcojavier@um.es
authors.forename: Francisco Javier
authors.initials: FJ
authors.lastname: Rodríguez-Lozano
authors.forename: C L
authors.initials: CL
authors.lastname: Insausti
authors.forename: L
authors.initials: L
authors.lastname: Meseguer
authors.forename: M C
authors.initials: MC
authors.lastname: Ramírez
authors.forename: S
authors.initials: S
authors.lastname: Martínez
authors.forename: J M
authors.initials: JM
authors.lastname: Moraleda
mesh_terms: D000328:Adult; D002453:Cell Cycle; D002454:Cell Differentiation; D003782:Dental Pulp / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D008964:Molar, Third; D010012:Osteogenesis / Q000502:physiology*; D010641:Phenotype; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology*