染色及激光显微切割对小鼠牙胚组织总RNA完整度的影响  

The effect of staining and LMD process on the integrity of total RNA of the tissues dissected from murine dental germ

作　　者：党平[1] 聂敏媛[1] 崔三哲[1] 汤楚华[1] 史亮[1] 郑翰圣[2] 董蕊[3] 施生根[1] 

机构地区：[1]解放军第306医院口腔科,北京100101 [2]韩国延世大学口腔医学院 [3]首都医科大学口腔医学院.口腔医学研究所

出　　处：《实用口腔医学杂志》2013年第3期319-322,共4页Journal of Practical Stomatology

基　　金：全军十一五课题重点项目(编号:06MA004)

摘　　要：目的:观察以常规染色结合激光显微切割(laser microdissection,LMD)技术精确分离获取小鼠牙胚间充质组织对所获得组织总RNA完整度的影响。方法:以LMD技术获取胚胎17 d(embryonic 17,E17)和出生后2 d(postnatal 2,PN2)两时期CD1小鼠下颌第一磨牙的牙囊和牙乳头组织;提取组织总RNA后再以2100生物分析仪所提供的核糖体RNA 28s/18s比值(28s/18s Ratio)、RNA完整度数值(RNA integrity number,RIN),以及样品电泳图的图形来综合判断样品完整度。结果:与未经LMD切割的对照组相比,经染色及LMD切割的实验组所提取牙胚组织样品总RNA完整度明显降低;非染色仅实行LMD组样品总RNA完整度高于染色及LMD组,但其完整度仍低于通常基因芯片实验要求。结论:常规染色及LMD过程对所提取牙胚组织总RNA完整度有显著影响。Objective： To investigate the effect of isolating the mesenchyma tissue of tooth germ by laser microdissection（LMD） and normal dyeing on RNA Integrity. Methods： The dental follicle and dental papilla tissues of murine first lower molar at El7 and PN2 stages were dissected by LMD6000 system. The integrity and quality check of RNA samples were examined by Bioanalyzer 2100 Sys- tem Agilent. The 28s/18s Ratio of ribosome RNA, RNA integrity number （RIN） and the peak pattern of the electrophoretogram were integrated as the index of RNA integrity. Results： The mesenehyma tissues of the first lower molar were successfully isolated by LMD system with or without normal dyeing. LMD cutting and dying processes showed significant effect on RNA integrity of the isola- ted tissues. Conclusion ： LMD may damage RNA integrity of dental germ tissues.

关 键 词：激光显微切割技术 牙胚发育 间充质 RNA完整度 

分 类 号：R78[医药卫生—口腔医学]