Establishment of cementblast cell lines and periodontal ligament cell lines

本研究旨在建立牙周膜成纤维细胞和成牙骨质细胞的细胞系，为牙周膜细胞的增殖和分化研究奠定基础。首先，我们开发了连续酶消化法分离区域细胞群（成牙骨质细胞，前成牙骨质细胞，PDL成纤维细胞和未分化细胞）从大鼠磨牙PDL证明PDL组成细胞的异质性。接下来，我们将永生相关基因如SV-40 T和hTERT基因转染到细胞群中以建立它们的细胞系.采用连续酶消化法从大鼠和人PDL中分离出具有不同生长潜能和分化能力的细胞群.通过SV-40 T转染大鼠磨牙牙周膜细胞亚群，获得牙骨质母细胞系和牙周膜成纤维细胞系.将hTERT基因转染到区域细胞群中，建立人成牙骨质细胞系和牙周膜成纤维细胞系.本研究通过SV-40 T和hTERT基因转染人牙周膜成纤维细胞，建立了人牙周膜成纤维细胞系和人牙周膜成纤维细胞系，可广泛应用于牙周膜成纤维细胞的研究。这些细胞系有望成为研究牙周膜细胞分化增殖调控机制的新型体外细胞模型，并可用于牙周组织再生、牙种植和牙移植等相关研究。

The aim of the study was to establish the cell lines of periodontal ligament (PDL) fibroblasts and cementoblasts for elucidating cellular proliferation and differentiation of PDL. First, we developed the sequential enzymatic digestion method for isolation of regional cell populations (cementoblasts, precementoblast, PDL fibroblasts and undifferentiated cells) from rat molar PDL to demonstrate the heterogeneity of PDL constitutive cells. Next, we transfected immortality-related genes such as SV-40T and hTERT genes into the cell populations to establish their cell lines.1. We isolated cell populations with different growth potential and differentiation from rat and human PDL by sequential enzymatic digestion.2. Cementblast cell lines and PDL fibroblast cell lines were obtained from rat molar PDL cell subpopulation by the SV-40T transfection.3. We established human cementoblast cell lines and PDL fibroblast cell lines by the hTERT gene transfection into the regional cell populations.4. Human PDL cell lines were established from a human cementifying fibroma by the SV-40T and hTERT genes transfection.We established human and rat cementoblast cell lines and PDL fibroblast cell lines that can be widely provided for researchers. It is expected that these cell lines can be the novel in vitro cell model to study the regulatory mechanism of differentiation and proliferation of PDL and related studies including periodontal tissue regeneration, dental implant and tooth transplantation.