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Title of Journal: Cell Tissue Res

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Abbravation: Cell and Tissue Research

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Springer-Verlag

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DOI

10.1007/s00425-005-0075-1

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1432-0878

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Establishing and characterizing human periodontal

Authors: Shinsuke Fujii Hidefumi Maeda Naohisa Wada Yoshio Kano Akifumi Akamine
Publish Date: 2006/01/12
Volume: 324, Issue: 1, Pages: 117-125
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Abstract

The periodontal ligament PDL is a highly specialized tissue connecting the cementum with the tooth socket bone and affects the life span of the tooth However little is known about the precise characteristics and regenerative mechanism of PDL cells because of the absence of specific markers and cell lines Therefore we aimed to establish three immortalized human PDL fibroblast cell lines by using simian virus40 Tantigen SV40TAg and human telomerase reverse transcriptase hTERT transfection expecting these cells to have the characteristics of primary cells The transfected cells were named STPLF The expression of SV40TAg and hTERT in all STPLF lines was verified by using the semiquantitative reverse transcriptionpolymerase chain reaction RTPCR method stretch PCR analysis or Western blotting analysis All STPLF showed stable proliferation at more than 120 population doublings PD whereas primary human PDL fibroblasts HPLF stopped at 10–20 PD Characterization by RTPCR analysis revealed that all STPLF genes mimicked the expression of their respective original HPLF genes STPLF expressed runtrelated transcription factor2 osterix alkaline phosphatase osteopontin osteocalcin periostin receptor activator of NFkappa B ligand osteoprotegerin epidermal growth factor receptor alphasmooth muscle actin and type XII collagen STPLF stimulated with 50 μg/ml ascorbic acid and 2 mM βglycerophosphate for 4 weeks produced more calcified deposits than did HPLF cultured with the same reagents These results suggest that each STPLF line retained the characteristics of the respective original HPLF that STPLF gained increased calcification activity and that STPLF are helpful tools for studying the biology and regenerative mechanisms of human PDL

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