永生化成釉细胞瘤细胞株TAM-1的建立

目的：建立永生化的成釉细胞瘤细胞株。方法：应用基因转染方法将SV40Tag整合到体外培养的成釉细胞瘤细胞基因组中，用PCR和RT－PCR检测SV40Tag的表达，单克隆培养筛选出永生化的成釉细胞瘤细胞株。结果：未转染细胞体外存活51d。转染SV40Tag的肿瘤细胞的寿命延长，克隆细胞生长良好，呈小多边形，传代25次后仍保持生长旺盛，命名为TAM－1。结论：SV40Tag可整合到釉细胞瘤细胞基因组中，并获得稳定表达，转染SV40Tag的成釉细胞 瘤细胞株TAM－1获得了永生性。

Establishment of immortalized ameloblastoma cell line TAM-1

Objective To establish an immortalized ameloblastoma cell line. Methods The primary cultured ameloblastoma cells were transfected with pRSV Tag using Transfect AMINE kit. Tansfected cells were passaged to pass through crisis period and immortalise. Results Cultured ameloblastoma cells were composed predominantly of closely packed small polygonal cells with epithelial morphology. They had limited life span of 51 days in vitro. The small polygonal cells were eventually replaced by large flattened cells and subsequently became senescent and dead. On the other side, those tumor cells transfected with SV40Tag could live for a longer time. The majority of them died in crisis period while the survived cells from crisis period gained the ability to proliferate. There was no morphological change in TAM 1 compared with original cultured cells. A cell clone was harvested which was alive and keeping on proliferating after having been subcultured for 25 times. It was named TAM 1. The epithelial origin of TAM 1 was confirmed by strong immunoreactivity for cytokeratin in contrast to negative vimentin expression. It was detected that SV40Tag had been transfected into TAM 1 genesome and expressed continuously by PCR and RT PCR. Conclusions TAM 1 is immortalized ameloblastoma cell line in vitro.