慢病毒介导的外源基因在人牙髓干细胞中的表达

牙髓干细胞（dental pulp stem cells,hDPSC）是牙源性的间充质干细胞,具有多向分化潜能.已有的研究表明,一些蛋白因子能够诱导牙髓干细胞的牙向分化,但尚无通过过量表达某些关键基因来诱导牙髓干细胞牙向分化的报道.利用绿色荧光蛋白作为报告基因,探究慢病毒介导的外源基因在牙髓干细胞中的表达,分离并鉴定人的恒牙牙髓干细胞,用携带绿色荧光蛋白标记的慢病毒原液感染DPSC,感染病毒后的DPSC进行传代以验证外源基因的稳定表达,并将感染慢病毒后的DPSC与羟基磷灰石/磷酸三钙（hydroxyapatite/tricalcium phosphate,HA/TCP）混合移植到小鼠肾囊膜下培养8周.结果表明：用绿色荧光蛋白标记的慢病毒能够整合到牙髓干细胞的基因组中并获得稳定的表达,感染慢病毒前后的牙髓干细胞增殖率没有发生改变,感染病毒的hDPSC与HA/TCP混合移植到肾囊膜下仍能够产生牙本质牙髓样结构,因此利用慢病毒载体介导的绿色荧光蛋白并不影响牙髓干细胞的生物学特性,说明在进一步利用慢病毒载体研究过量表达基因在牙髓干细胞牙向分化的作用中,绿色荧光蛋白可以作为标记蛋白.

Expression of exogenous in the dental pulp stem cells mediated by lentiviral vector

To isolate,culture and identify human dental pulp stem cells（hDPSCs） and investigate the expression of exogenous in hDPSCs.DPSCs were transduced with EGFP by lentiviral vector.The effect of infection on proliferation of DPSCs was evaluated by MTT.Approximately 5×106 of DPSCs （third passage） were mixed with 40 mg of HA/TCP ceramic powder and then transplanted s.c.into the dorsal surface of 10-week-old immunocompromised beige mice.Results indicates： EGFP was stably introduced to DPSCs by lentiviral vector.The infection had no significant effect on the proliferation of DPSCs.After 8 weeks posttransplantation,DPSCs which infected EGFP generated a dentin-like structure lining the surfaces of the HA/TCP particles.Exogenous can be stably expressed in hDPSC mediated by lentiviral vector.And EGFP can be use for labeling gene in the lentiviral vector.