用基因转染技术建立人成牙本质细胞株

The odontoblast is thought to be responsible for dentin formation and mineralization during tooth development and dentin reparation. However, the precise mechanism of odontogenic differentiation and dentinogenisis are often hampered by the limit life span of primary cells that undergo senescence after about 10 passages. Thus, the purpose of this research was to establish and preliminarily characterize an immortalized human odontoblast-like cell line by transfection with human telomerase reverse transcriptase (hTERT) gene. Primary human odontoblast-like cells were cultured and transfected with plasmid pCIneo-hTERT by LipofectAMINE 2000 reagent. Under selection with G418 for a month, five drug-resistant cell clones gradually formed and were selected using filter paper and expanded for continuous culture in vitro. Cells derived from clone 5b had been maintained in culture for about 6 months, up to 56 passages, which was designated as hTERT-immortalized human odontoblast-like cell line (hTERT-hOd-l). hTERT-hOd-l cells were almost normal without significant transformed phenotype, and remained the biological characteristics of primary cells, and could synthesize dentin extracellular matrix (DECM) protein to form mineralized matrix in vitro. Our results suggest that transfection of human odontoblast-like cells with hTERT resulted in an immortalized cell line hTERT-hOd-l maintaining phenotypic characteristics of native odontoblast. This cell line would facilitate further studies on odontoblasts, dentinogenesis and tissue engineering of dentin..Keywords: odontoblast; immortalization; cell line; hTERT; dentin

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