人成牙本质细胞样细胞的原代培养

Primary culture of human odontoblast-like cell

目的 :培养人原代成牙本质细胞。方法 :取引产的 8月龄死胎 ,剥离乳磨牙胚牙乳头 ,组织块法培养。然后采用滤纸片法挑取了 3个与成牙本质细胞形态相似的细胞克隆 ,扩大培养。对培养细胞从形态学、矿化结节、碱性磷酸酶 (alkalinephosphatase,ALP)活性、人牙本质基质蛋白 - 1(humandentinmatrixprotein- 1,hDMP - 1)和人牙本质涎磷蛋白 (humandentinsialophosphoprotein ,hDSPP)在mRNA水平上的表达等方面进行鉴定。结果 :有一个克隆来源的细胞呈梭形、有单侧较长细胞突起 ,未见核极化 ,同时它们具矿化功能 ,表达人成牙本质细胞特异性标志hDMP - 1、hDSPPmRNA。结论 :该培养细胞为人成牙本质细胞样细胞 ,为进一步的有关研究奠定了基础。

AIM:To cultivate primary human odontoblast. METHODS: Human deciduous molar tooth germs were isolated from a still-birth, 8 month old female foetus. The dental papillae were mechanically removed and cells were prepared by using tissue explant culture technique. These cell clones, which took on similar morphology with odontoblast, were selected by using filter paper and expanded in culture. The primary cultured cells were identified by morphology, formation of mineralized nodule, activity of alkaline phosphatase (ALP), and mRNA expression of huamn dentin matrix protein-1(hDMP-1)and human dentin sialophosphoprotein (hDSPP).RESULTS:The cells from one of the three clones took on a fibroblast-like form with single and long process, without polarized nucleus. Also, the cells had the ability of mineralization and expressed specific markers of odontoblast, hDMP-1 and hDSPP at mRNA level. CONCLUSION: The cultured cells are human odontoblast-like cells, which would facilitate further studies.