Department of Pediatric Dentistry, School of Dentistry, Iwate Medical University, Morioka 020-8505, Japan.
Int J Mol Med. 2010 Nov;26(5):701-5. doi: 10.3892/ijmm_00000516.
Although periodontal ligament (PDL) cells have previously been isolated from permanent teeth, they have not been isolated from deciduous teeth. Here, we used human telomerase reverse transcriptase (hTERT) induction to establish the first immortalized PDL cell lines derived from deciduous teeth. Cells were transfected with plasmids containing hTERT. Single-cell cloning was then performed using the limited dilution method. Reverse transcriptase polymerase chain reaction and stretch PCR were used to detect hTERT expression in the clones. In order to determine whether the clones could differentiate into osteoblasts, we stimulated the cells with ascorbic acid and β-glycerophosphate. We success-fully 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 hTERT 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 SH 9 cell line differentiated into osteoblastic cells. This is the first report of the immortalization of PDL cells derived from deciduous teeth. These cells could be useful in studies investigating the cellular mechanisms and regenerative processes of human PDL cells.
虽然牙周膜（PDL）细胞以前曾从恒牙中分离出来，但尚未从乳牙中分离出来。在这里，我们使用人端粒酶逆转录酶（hTERT）诱导来建立第一个从乳牙中分离出来的永生化 PDL 细胞系。将含有 hTERT 的质粒转染细胞。然后使用有限稀释法进行单细胞克隆。使用逆转录聚合酶链反应和延伸 PCR 检测克隆中的 hTERT 表达。为了确定克隆是否可以分化为成骨细胞，我们用抗坏血酸和β-甘油磷酸刺激细胞。我们成功获得了 3 个单细胞克隆，并将它们命名为来自人类乳牙 PDL 的单细胞（SH）9、10 和 11。所有 SH 细胞均表现出 hTERT 表达和>80 个群体倍增后的稳定增殖，并表达 PDL 细胞的标记基因，包括 Scleraxis、periostin、cementum-derived protein 23 和 tenomodulin。尽管所有克隆均表达成骨细胞标记物，但只有来自 SH 9 细胞系的克隆分化为成骨细胞。这是乳牙来源的 PDL 细胞永生化的首次报道。这些细胞可用于研究人 PDL 细胞的细胞机制和再生过程。
