Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
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In vitro characterization of human dental pulp stem cells isolated by three different methods

  • Jang, Ji-Hyun (Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong) ;
  • Lee, Hyeon-Woo (Department of Pharmacology, School of Dentistry) ;
  • Cho, Kyu Min (Department of Conservative Dentistry, Graduate School, Kyung Hee University) ;
  • Shin, Hee-Woong (School of Dentistry, University of Western Australia) ;
  • Kang, Mo Kwan (School of Dentistry and Jonsson Comprehensive Cancer Center, UCLA) ;
  • Park, Sang Hyuk (Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong) ;
  • Kim, Euiseong (Microscope Center, Department of Conservative Dentistry and Oral Science Research Center, College of Dentistry, Yonsei University)
  • Received : 2016.06.17
  • Accepted : 2016.08.08
  • Published : 2016.11.30
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Abstract

Objectives: In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods: HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results: Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions: Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

Keywords

References

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