McLachlan Julia L, Smith Anthony J, Cooper Paul R
Oral Biology, School of Dentistry, University of Birmingham, St. Chads Queensway, Birmingham B4 6NN, UK.
Arch Oral Biol. 2003 Oct;48(10):731-6. doi: 10.1016/s0003-9969(03)00152-3.
Isolation of sufficient quantities of pure populations of odontoblasts from healthy and diseased teeth will facilitate our understanding of dentinogenesis during development and repair. Here we describe a novel Piezo-power microdissection (PPMD) technique for the isolation of pure populations of odontoblasts and pulpal tissue from formalin-fixed, paraffin-embedded, mature, healthy and carious human teeth. Odontoblasts and pulpal tissue gene expression were subsequently studied in ribonucleic acid isolated from PPMD preparations using a semi-quantitative reverse transcription polymerase chain reaction approach. Data confirmed that the genes for dentine sialophosphoprotein and Nestin are preferentially expressed in odontoblasts, whilst the genes for both collagen-1alpha and collagen-3alpha were expressed preferentially in pulpal tissue, particularly in carious samples. PPMD provides a novel and powerful approach to isolate pure populations of dental tissues and cells from fixed specimens for subsequent downstream molecular analyses.
从健康和患病牙齿中分离出足够数量的纯成牙本质细胞群体,将有助于我们了解发育和修复过程中的牙本质形成。在此,我们描述了一种新型的压电动力显微切割(PPMD)技术,用于从福尔马林固定、石蜡包埋、成熟、健康和龋坏的人类牙齿中分离出纯成牙本质细胞群体和牙髓组织。随后,使用半定量逆转录聚合酶链反应方法,对从PPMD制备物中分离的核糖核酸中的成牙本质细胞和牙髓组织基因表达进行了研究。数据证实,牙本质涎磷蛋白和巢蛋白基因在成牙本质细胞中优先表达,而Ⅰ型胶原和Ⅲ型胶原基因则优先在牙髓组织中表达,尤其是在龋坏样本中。PPMD提供了一种新颖且强大的方法,可从固定标本中分离出纯的牙齿组织和细胞群体,用于后续的下游分子分析。