Profiles of gene expression on periodontal ligament cells

为了确定牙周膜细胞上特异性基因的表达，我们对单细胞来源的克隆细胞进行了刺激后基因表达的变化分析。本实验采用常规培养方法，从单细胞来源，制备了有限数量的人牙周膜细胞。然后，我们不得不在培养基中加入几种生长因子，以维持长期培养。我们分析了成纤维细胞生长因子-2（FGF-2）对人牙周细胞增殖和复制能力的影响。添加FGF-2可使细胞增殖增加10 ~ 15%。FGF-2还增强了有限稀释法中培养细胞的集落形成能力。这些影响在年轻和年老的细胞之间没有改变，并且在培养细胞的整个寿命期间都是如此。我们分析了FGF-2对连续长期细胞培养中细胞复制能力的影响。FGF-2的加入促进了生命早期的增殖，但在生命晚期却抑制了增殖。总的来说，FGF-2的加入导致复制能力下降。用半定量RT-PCR方法检测了p53和p16基因在不同寿命期的培养细胞中的表达水平，结果表明，在不同寿命期的培养细胞中，p53和p16基因的表达水平无显著性差异。这些结果表明，FGF-2，这是已知的几种细胞的有效的有丝分裂原，在长期培养中具有逆转作用。

In order to determine specific gene expression on periodontal ligament cells, we analyzed the change of gene expression after stimulation against the cloned cells that were derived from single-cell. We only prepared limited amount of human periodontal ligament cells which were derived from single-cell by using popular media. Then we had to add several growth factors,into media to maintain long term-culture. We analyzed the effects of fibroblast growth factor-2 (FGF-2) on proliferation and replicative capacity in human periodontal cells. Addition of FGF-2 on logarithmically growing cell enhanced proliferation of cell number by 10 to 15 %. FGF-2 also enhanced colony forming capacities on cultured cells in the limited dilution methods. These effects did not change between young and old cells, and suggested same throughout the life-span of cultured cells. We analyzed the effect on FGF-2 to replicative capacity in continuous long-term cell culture. Addition of FGF-2 enhanced proliferation in early stage of life-span, however, it caused inhibition of proliferation at late stage of life span. Totally, addition of FGF-2 caused decreased replicative capacity. We also determined gene expression levels of p53 and p16 into the cultured cells at different stage of life-span by semi-quantified RT-PCR method, and identified no significant differences into the cells. These results suggest that FGF-2, which was known a potent mitogen for several kind of cells, has reverse effect during long-term culture.