title: Immortalization and characterization of human dental mesenchymal cells.
abstract: OBJECTIVE
  Due to the rarity of human embryonic samples and limited proliferating capability of primary human dental mesenchymal cells, it is valuable to create an immortalized human dental mesenchymal cell line for studying dental mesenchymal cell differentiation and signalling pathways during dentinogenesis in humans.
  
  
  METHODS
  In this study, dental mesenchymal cells from human molar tooth germs at 19-week gestation were isolated and immortalized with pSV40. Single cell colonies were then selected by 96-well plate dilution. The immortalized cell line was characterized using immunofluorescent microscopy, RT-PCR and Western blot for the expression of SV40 large T antigen and five genes specific for the mesenchymal stage during tooth development. The differentiation and mineralization activities of the immortalized and primary cells were compared using adipogenic and calcifying induction.
  
  
  RESULTS
  The immortalized dental mesenchymal cell line displayed a higher proliferation rate, expressed several tooth-specific markers including Msx1, Pax9, Lhx6, Barx1, and Runx2, and maintained the ability to differentiate and form mineralized nodules.
  
  
  CONCLUSIONS
  Our results demonstrated that the immortalized human mesenchymal cell line retained the characteristics similar to primary human dental mesenchymal cells and can be used for studying the mechanisms of human dental mesenchymal cell differentiation and signalling pathways involved in human odontogenesis.
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China. Electronic address: ydhuang@fjnu.edu.cn.
authors: Yide
authors: Y
authors: Huang
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China.
authors: Yun
authors: Y
authors: Yang
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China.
authors: Meiqin
authors: M
authors: Jiang
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China.
authors: Meizhen
authors: M
authors: Lin
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China.
authors: Shuiqin
authors: S
authors: Li
authors: College of Life Sciences, Fujian Normal University, Fujian Key Laboratory of Developmental and Neural Biology, Fujian, PR China. Electronic address: yaolin@fjnu.edu.cn.
authors: Yao
authors: Y
authors: Lin
mesh_terms: D037881:Aborted Fetus / Q000166:cytology; D002113:Calcification, Physiologic; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008648:Mesoderm / Q000166:cytology*; D008963:Molar / Q000166:cytology; D014083:Tooth Germ / Q000166:cytology* / Q000378:metabolism
keywords: Dental mesenchymal cell; Immortalization; SV40 large T antigen; Tooth-specific marker