In vitro characterization of human dental pulp cells: various isolation methods and culturing environments

  • George T.-J. HuangEmail author
  • Wataru Sonoyama
  • James Chen
  • Sang Hyuk Park
Regular Article


Our purpose was to characterize human dental pulp cells isolated by various methods and to examine the behavior of cells grown under various conditions for the purpose of pulp/dentin tissue engineering and regeneration. We compared the growth of human pulp cells isolated by either enzyme digestion or the outgrowth method. Expression of dentin sialophosphoprotein, Cbfa1, and two types of collagen (I and III) in these cells was examined by Western blot or reverse transcription/polymerase chain reaction. Growth of pulp cells on dentin and in collagen gel was also characterized. We found that different isolation methods give rise to different populations or lineages of pulp cells during in vitro passage based on their collagen gene expression patterns. Cells isolated by enzymedigestion had a higher proliferation rate than those isolated by outgrowth. Pulp cells did not proliferate or grew minimally on chemically and mechanically treated dentin surface and appeared to establish an odontoblast-like morphology with a cytoplasmic process extending into a dentinal tubule as revealed by scanning electron microscopy. The contraction of the collagen matrix caused by pulp cells was dramatic: down to 34% on day 14. Our data indicate that (1) the choice of the pulp cell isolation method may affect the distribution of the obtained cell populations, (2) a treated dentin surface might still promote odontoblast differentiation, and (3) a collagen matrix may not be a suitable scaffold for pulp tissue regeneration because of the marked contraction caused by pulp cells in the matrix. The present study thus provides important information and a basis for further investigations pre-requisite to establishing pulp tissue engineering/regeneration protocols.


Dental pulp cells Dentin sialophosphoprotein Cbfa1 Collagen type I/III Reverse transcription/polymerase chain reaction Scanning electron microscopy Human 



We greatly appreciated the technical assistance of Alicia Thompson (Center for Electron Microscopy and Microanalysis) with the SEM analysis.


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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • George T.-J. Huang
    • 1
    • 2
    • 3
    • 5
    Email author
  • Wataru Sonoyama
    • 4
  • James Chen
    • 1
    • 6
  • Sang Hyuk Park
    • 1
  1. 1.Division of Associated Clinical Specialties, Section of EndodonticsUCLA School of DentistryLos AngelesUSA
  2. 2.Division of Oral Biology and Medicine, and Orofacial PainUCLA School of DentistryLos AngelesUSA
  3. 3.Dental and Craniofacial Research InstituteUCLA School of DentistryLos AngelesUSA
  4. 4.Department of Oral and Maxillofacial RehabilitationOkayama University Graduate School of Medicine, Dentistry, and Pharmaceutical SciencesOkayamaJapan
  5. 5.College of Dental Surgery, Dental School, Department of Endodontics, Prosthodontics, and Operative DentistryUniversity of MarylandBaltimoreUSA
  6. 6.Chino HillUSA

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