小鼠成牙本质样细胞的原代培养及鉴定

Primary culture and identification of mouse odontoblast-like cells

目的:原代培养小鼠成牙本质细胞,为诱导胚胎干细胞(ES细胞)向成牙本质细胞分化提供基础。方法:取材于出生1周昆明小鼠的下颌切牙牙胚,分离牙乳头组织,采用消化法对小鼠牙乳头细胞进行分离培养;用细胞平板克隆技术,挑选具有成牙本质细胞形态特征的细胞克隆,扩大培养;并从光镜观察、电镜观察和小鼠牙本质涎磷蛋白(dentinsialophoprotein,DSPP)在mRNA水平上的表达等方面进行鉴定。结果:培养细胞胞质丰富,形态呈梭形,具有较长的细胞突起。超微结构观察,细胞富含高尔基复合体、核糖体和粗面内质网,并特异性表达DSPPmRNA。结论:该研究成功培养出小鼠成牙本质样细胞,该方法可用于成牙本质细胞的体外研究。

PURPOSE： To culture primary mouse odontoblast and to provide a base for study on inducing ES cells to odontoblast. METHODS： Lower incisor germs were removed from 1-week-old mouse. The dental papillae were isolated in microscope, and the dental papillae cells were dispersed using 0.25% collagenase and 0.25% trypsin. The cell clones, which had similar morphology to odontoblasts, were selected for further culturing. The primary cultured cells were identified by light and electron microscopes and mRNA expression of mouse dentin sialophoprotein. RESULTS： The cultured cells had the same morphology and ultrastructure. They were rich in Golgi＇s complex, ribosome and rough endoplasmic reticulum. These cells expressed DSPP at mRNA levels. CONCLUSION： The cultured cells were mouse odontoblast-like cells. The method could be used for the study of odontal cells in vitro. Supported by Science and Technology Development Fund of Shanghai Municipality （Grant No.03ZR14027）.