title: In vitrocharacterization of human dental pulp stem cells isolated by three different methods.
abstract: OBJECTIVE
  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.
  
  
  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 vitrodifferentiation 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.
authors.affiliation: Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, Seoul, Korea.
authors.forename: Ji-Hyun
authors.identifier: 0000-0002-4672-3381
authors.initials: JH
authors.lastname: Jang
authors.affiliation: Department of Pharmacology, School of Dentistry, Kyung Hee University, Seoul, Korea.; Oral Biology Research Institute, School of Dentistry, Kyung Hee University, Seoul, Korea.
authors.forename: Hyeon-Woo
authors.initials: HW
authors.lastname: Lee
authors.affiliation: Department of Conservative Dentistry, Graduate School, Kyung Hee University, Seoul, Korea.
authors.forename: Kyu Min
authors.initials: KM
authors.lastname: Cho
authors.affiliation: School of Dentistry, University of Western Australia, Nedlands, WA, Australia.
authors.forename: Hee-Woong
authors.initials: HW
authors.lastname: Shin
authors.affiliation: School of Dentistry and Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA, USA.
authors.forename: Mo Kwan
authors.initials: MK
authors.lastname: Kang
authors.affiliation: 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.
authors.forename: Sang Hyuk
authors.identifier: 0000-0002-0518-6938
authors.initials: SH
authors.lastname: Park
authors.affiliation: Microscope Center, Department of Conservative Dentistry and Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Euiseong
authors.identifier: 0000-0003-2126-4761
authors.initials: E
authors.lastname: Kim
keywords: Dental pulp stem cells; Isolation method; Mesenchymal stem cells