Notch信号分子于小鼠牙髓干细胞样细胞表达的研究
Characterization of Notch Gene Involved in Genetic Regulatory Networks in Dental Pulp Stem Cells
作者：陆群,吴补领,王冀姝,韩骅,周学东
Author：LUQun1,WUBu- ling2,WANG Ji-shu3,HANHua3,ZHOUXue-dong1
收稿日期：2004-02-25 年卷（期）页码：2004,22(01):54-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：牙髓干细胞,Notch,牙齿发育,反转录聚合酶联反应,
Key words：dental pulp stem cells,Notch,tooth differentiation,reverse transcription-PCR,
基金项目：
国家973计划资助项目子项目(2001CB509906);全国高等学校骨干教师计划资助(1999年)
中文摘要
目的 研究Notch基因在小鼠牙髓干细胞样细胞表达。方法 采用酶消化培养法获得小鼠的单个牙髓细胞悬液,调整细胞密度为1×104个/孔细胞,干细胞培养液培养14 d,挑选细胞克隆扩增,提取细胞的总RNA,反转录聚合酶联反应(RT-PCR)检测Notch基因的表达。结果 小鼠牙髓细胞呈集落状生长,克隆形成率约为1·6 ~2·5 个/104细胞,所形成的集落细胞结合紧密,细胞胞体小、胞核大,RT-PCR结果显示Notch的mRNA在牙髓干细胞样细胞中有表达。结论 培养的集落状生长的小鼠牙髓细胞具有干细胞增殖快的特性,Notch基因于牙髓干细胞中表达,表明Notch信号参与了牙髓干细胞样细胞的早期分化的调控过程。
英文摘要
Objective To investigate the characterization of Notch gene involved in genetic regulatory networks in dental pulp stem cells.Methods The pulp tissue was separated frommouse teeth and digested by collagenase type I. Single-cell suspensions of dental pulp were seeded into 6-well plates with alpha modification of Eagle′s medium supplemented with ES cell qualified Fetal Bovine Serum. Colony-forming efficiency was assessed in 14ds culture. Transcripts for Notch were detected by reverse transcrip- tion-PCR by using total RNA isolated from cells.Results There were clonogenic cells in dental pulp cell and the incidence of colony-forming cells derived from mouse dental pulp cells was 1·6~2·5 colonies/104plate. Mouse-specific Notch mRNA ex- pressed in colony-forming cells.Conclusion Notch mRNA expressing in colony-forming cells provided a more detailed under- standing of mouse dental pulp stem cell biology.
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