乳歯歯根膜細胞による再生医学研究の可能性の検討  [in Japanese] Investigation of New Strategy for Periodontal Tissue Regeneration Using with Human Immortalized Periodontal Ligament Cell Line Derived from Deciduous Teeth  [in Japanese]

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Abstract

脱落し廃棄される乳歯の歯髄や歯根膜に多分化能を持つ細胞の存在が報告されている。小児歯科医療の現場において患者自身から採取され廃棄される抜去歯から細胞を分離し利用可能であれば,倫理的・免疫学的・安全性の問題を回避でき,さらに経済的にも安価な再生医療の資源になり得ると考えられる。しかし乳歯歯根膜由来の細胞には不死化細胞株が存在せず,そのため十分な再生医学研究が行われていない。そこで再生医学研究の実験材料を作る目的で,ヒト乳歯歯根膜細胞へテロメラーゼ遺伝子の導入により不死化細胞株の樹立を行った。single cell derived from human deciduous PDL(SH)と名付けSH 9, 10, 11 の3 株を分離した。全ての細胞株において,集団倍化指数(PDs)は80 PDs を越え,ヘイフリックの限界の50 PDs を越えて増殖していることから不死化していると考えられた。全ての細胞株で歯根膜のマーカーの遺伝子を発現していた。また骨芽細胞への分化能について検討した結果,SH 9 株のみが石灰化結節を形成した。さらにSH 9 株は幹細胞の遊走誘導をコントロールするstromal cell-derived factor-1 <i>α </i>を発現し,歯根膜組織の再生および恒常性の維持に働いていることが示唆された。以上のことから乳歯歯根膜細胞の不死化細胞株は,今後の再生医学研究に非常に有用であると考えられた。

Recently, several studies have demonstrated that periodontal ligament (PDL) cells differentiate into cementoblastic cells and adipogenic cells <i>in vitro. </i>Therefore, the PDL probably contains pluripotent progenitor cells or putative stem cells. Until now, bone-marrow-derived MSCs (BMMSCs) are good candidates for the treatment of mesenchymal tissue as a source of cells for cell-based therapeutic strategies. Although BMMSCs are very usuful for clinical applications, bone marrow aspiration for MSCs extraction is an invasive and painful procedure for the donor. In addition, the number, proliferation and differentiation potential of BMMSCs decline with increasing age. However, isolating MSCs derived from PDL tissue of deciduous tooth can be non-invasive and economical procedure when permanent tooth replace deciduous tooth.Therefore, it has been thought that PDL cell line from deciduous teeth could be useful tool for the investigation of regenerative medicine. Nevertheless, clonal PDL cell lines have not been established from deciduous teeth.Here, we used human telomerase reverse transcriptase (<i>hTERT</i>) induction to establish the first immortalized PDL cell lines derived from deciduous teeth. We successfully obtained 3 single-cell clones, and named them single cell derived from human deciduous PDL (SH) 9, 10 and 11. All the SH cells showed <i>hTERT </i>expression and stable proliferation after >80 population doublings and expressed the marker genes of PDL cells, including scleraxis, periostin, cementum-derived protein 23,and tenomodulin. Although all the clones expressed osteoblastic markers, only the clones from the SH9 cell line differentiated into mature osteoblastic cells which could form calcified nodules <i>in vitro. </i>Recent study have showed that stromal cell-derived factor (SDF)-1 <i>α </i>play a crucial role in stem cell homing and recruitment to injured sites. However, no information is available about SDF-1 <i>α </i>in periodontal tissues. The second aim of this study was to evaluate whether fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-<i>β</i>1 could affect SDF-1 <i>α </i>expression by immortalized PDL cells derived from deciduous tooth (SH 9 cells) <i>in vitro. </i>FGF-2 inhibited SDF-1 <i>α </i>expression at about 10% of control by 48 h, TGF-<i>β</i>1 induced it at about 6 fold of control between 6 h and 12 h after treatment. These results suggested that SDF-1 <i>α </i>produced by PDL might play an important role in adult stem cell and progenitor cell release, recruitment and homing to injury sites.This is the review of the immortalization of PDL cells derived from deciduous teeth and the regulation of SDF-1 <i>α </i>expression in PDL cells with FGF-2 and TGF-<i>β</i>1. These cells could be useful in studies investigating the cellular mechanisms and regenerative processes of human PDL cells.

Journal

  • The Japanese Journal of Pediatric Dentistry

    The Japanese Journal of Pediatric Dentistry 50(1), 7-14, 2012-03-25

    Japanese Society of Pediatric Dentistry

References:  38

Codes

  • NII Article ID (NAID)
    10030558370
  • NII NACSIS-CAT ID (NCID)
    AN00116228
  • Text Lang
    JPN
  • Article Type
    REV
  • ISSN
    05831199
  • Data Source
    CJP  J-STAGE 
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