In vitro characterization of human dental pulp stem cells isolated by three different methods.
10.5395/rde.2016.41.4.283
- Author:
Ji Hyun JANG
1
;
Hyeon Woo LEE
;
Kyu Min CHO
;
Hee Woong SHIN
;
Mo Kwan KANG
;
Sang Hyuk PARK
;
Euiseong KIM
Author Information
1. Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, Seoul, Korea. shpark94@khu.ac.kr
- Publication Type:Original Article
- Keywords:
Dental pulp stem cells;
Isolation method;
Mesenchymal stem cells
- MeSH:
Antigens, Differentiation;
Blotting, Western;
Dental Pulp*;
Digestion;
Humans*;
In Vitro Techniques*;
Mesenchymal Stromal Cells;
Methods*;
Polymerase Chain Reaction;
Stem Cells*;
Tissue Engineering
- From:Restorative Dentistry & Endodontics
2016;41(4):283-295
- CountryRepublic of Korea
- Language:English
-
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.
