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Immortalization and characterization of human dental papilla cells with odontoblastic differentiation
Author(s) -
Yang G. B.,
Li X. Y.,
Yuan G. H.,
Liu H.,
Fan M. W.
Publication year - 2013
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/iej.12029
Subject(s) - dental papilla , nestin , dmp1 , cell culture , microbiology and biotechnology , immortalised cell line , odontoblast , population , cellular differentiation , biology , transfection , stem cell , pathology , gene , genetics , neural stem cell , medicine , pulp (tooth) , environmental health , viral matrix protein
Aim To establish a cell line of immortalized human dental papilla cells ( hDPC s). Methodology Primary hDPC s were cultured and infected with lentivirus containing the hTERT gene. Integration and transcription of the hTERT gene were verified by PCR. The characteristics of the cells, such as morphology, proliferation and mineralization, were analysed. Also, the expression of odontoblastic‐related markers including ALP, DMP1, DLX3, OSX, DSP and Nestin, was detected by immunohistochemistry and real‐time RT‐PCR. Results hTERT gene was integrated into genomic DNA of immortalized cells ( hDPC ‐TERT) and transcribed into mRNA . With long‐time culture, hDPC ‐TERT bypassed senescence and grew over 120 population doublings. hDPC ‐TERT cells have a higher proliferation rate, but retain the phenotypic characteristics of the primary hDPC s, and so was ALP activity and mineralization activity. Furthermore, the hDPC ‐TERT cells express no DSP and Nestin with maintenance medium, but highly expressed DSP and Nestin after odontoblastic induction. Conclusions A line of immortalized human dental papilla cells, which remains in an undifferentiated state and has odontoblastic differentiation potential, was established. This cell line can be used as a cell model for studying the mechanism of the initiation of odontoblast differentiation.