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Restor Dent Endod. 2016 Nov;41(4):283-295. Epub 2016 Oct 12.

In vitro characterization of human dental pulp stem cells isolated by three different methods.

Author information

1
Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, Seoul, Korea.
2
Department of Pharmacology, School of Dentistry, Kyung Hee University, Seoul, Korea.; Oral Biology Research Institute, School of Dentistry, Kyung Hee University, Seoul, Korea.
3
Department of Conservative Dentistry, Graduate School, Kyung Hee University, Seoul, Korea.
4
School of Dentistry, University of Western Australia, Nedlands, WA, Australia.
5
School of Dentistry and Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA, USA.
6
Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, Seoul, Korea.; Oral Biology Research Institute, School of Dentistry, Kyung Hee University, Seoul, Korea.; Department of Conservative Dentistry, Graduate School, Kyung Hee University, Seoul, Korea.
7
Microscope Center, Department of Conservative Dentistry and Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Korea.

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:

Dental pulp stem cells; Isolation method; Mesenchymal stem cells

Conflict of interest statement

No potential conflict of interest relevant to this article was reported.

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