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Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling
Published on: January 18, 2015
Establishment of immortalized ameloblastoma cell line TAM-1
1Department of Oral and Maxillofacial Surgery, Guanghua Stomatological Faculty, Sun Yat-sen University, Guangzhou 510060, China.
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 immortalize.
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.
