Characterization of human dental pulp-derived cell lines
Characterization of human dental pulp-derived cell lines
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DOI：
10.1111/j.1365-2591.2008.01409.x
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发表时间：
2008-07-01
影响因子：
5
通讯作者：
Komiyama, K.
中科院分区：
文献类型：
--
作者：
Suguro, H.;Asano, M.;Komiyama, K.
目的建立不同类型牙髓成纤维细胞系，并对其进行鉴定。方法采用脂质体转染法将SV40大T抗原导入人牙髓组织成纤维细胞。筛选G418耐药细胞，采用有限稀释法建立不同的细胞系。用抗成纤维细胞抗体、抗波形蛋白抗体、抗I型胶原抗体和III型抗体对每个克隆进行免疫荧光染色，以确定细胞的谱系。结果通过转染SV40大T抗原，获得了9个不同的细胞克隆。所有细胞克隆均被抗成纤维细胞、抗波形蛋白、抗I型和III型胶原抗体染色阳性。通过差异显示实验，检测到8个不同的基因，这些基因在每个细胞克隆中的表达水平不同。经过测序和数据库搜索，发现一个基因与T细胞标记Thy-1相同。结论人牙髓成纤维细胞系具有不同的基因表达谱，提示细胞亚群的存在。
Aim To establish and characterize different types of fibroblastic cell lines derived from dental pulp tissue.Methodology Human dental pulp tissue-derived cells were transfected with SV40 large T antigen by Lipofectamine transfection method. Geneticin (G418)-resistant cells were selected and different cell lines were established by a limiting dilution method. To characterize the lineages of cells, each clone was immunofluorescently stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. Total RNA was extracted from each clone and subjected to a differential display experiment.Results By transfecting SV40 large T antigen, nine different cell clones were obtained. All these cell clones were positively stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. With differential display experiment, eight different genes, the expression levels of these genes were varied amongst each cell clone, were detected. After sequencing and database search, one gene was revealed to be identical to T-cell marker, Thy-1. Thy-1 expression in dental pulp tissue was confirmed by immunohistochemical staining.Conclusion Fibroblastic cell lines derived from human dental pulp tissue possessed different gene expression profiles suggesting the existence of subpopulations.