In vitro characterization of human dental pulp stem cells isolated by three different methods
장지현, 이현우, Cho Kyu-Min, 신희웅, Kang Mo-Kwan, 박상혁, 김의성,
소속 상세정보
장지현 ( Jang Ji-Hyun ) - Kyung Hee University Dental Hospital at Gangdong Department of Conservative Dentistry
이현우 ( Lee Hyeon-Woo ) - Kyung Hee University College of Medicine Department of Pharmacology
( Cho Kyu-Min ) - Kyung Hee University Graduate School Department of Conservative Dentistry
신희웅 ( Shin Hee-Woong ) - University of Western Australia School of Dentistry
( Kang Mo-Kwan ) - Jonsson Comprehensive Cancer Center School of Dentistry
박상혁 ( Park Sang-Hyuk ) - Kyung Hee University Dental Hospital at Gangdong Department of Conservative Dentistry
김의성 ( Kim Eui-Seong ) - Yonsei University College of Dentistry Department of Conservative Dentistry
KMID : 1034420160410040283
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.
키워드
Dental pulp stem cells; Isolation method; Mesenchymal stem cells
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