大鼠成牙骨质细胞的体外培养及其生物学特性的实验研究  

In vitro culture and biological characteristics of rat cementoblasts

作　　者：张斌[1] 李岩峰[2] 

机构地区：[1]锦州医学院附属一院口腔科,121001 [2]北京301医院口腔科

出　　处：《实用口腔医学杂志》2004年第3期307-311,共5页Journal of Practical Stomatology

摘　　要：目的 :探讨体外培养源于大鼠磨牙牙骨质的成牙骨质细胞 (cementoblast ,CMB)的可行性 ,以期建立能排除牙周其它细胞干扰的CMB体外实验模型。方法 :使用组织块结合酶消化的二步法 ,分别培养来自牙骨质的CMB ,来自牙槽骨的成骨细胞 (osteoblast ,OB)和来自牙周韧带 ( periodontalligament ,PDL)的成纤维细胞。然后对所培养的细胞分别进行骨钙素免疫组化染色和碱性磷酸酶 (ALP)活力测定 ,并利用茜素红S ,检测在条件培养基下细胞形成矿化结节的情况。结果 :3种牙周细胞都可从约 12周大鼠第一磨牙根周获得进行原代培养。骨钙素在体外培养的CMB和成骨细胞中被强烈表达 ,在PDL成纤维细胞 (PDLC)中几乎不表达。成骨细胞ALP活性最强 ,在成纤维细胞中适中 ,在CMB中几乎无活性。CMB形成矿化结节茜素红S染色阳性。结论 :CMB可以在体外成功培养 ,并可与牙槽骨OB、PDLC相区别。CMB在形态和蛋白表达等很多地方与成骨细胞相似 ,如表达矿化相关蛋白等 ,但它几乎没有ALP活性 ;矿化条件下 。Objective: To investigate the feasibility of culturing and identifying the cementoblasts in vitro and to study the biological characteristics of rat cementoblasts.Methods: Osteoblasts, PDL fibroblasts and cementoblasts were obtained from alveolar bone and the root surface of rat first molars and cultured with a 2-step method of enzyme digestion/explant technique. Bone osteocalcin(OCN) and alkaline phosphatase(ALP) were detected by quantitative immunohistochemical examination. Alizarin Red S was used to examine the formation of calcified nodules under conditional culture. Results: Primarily cultured cells could be subcultured. Osteoblasts were identified by the expression of OCN and ALP at heigh level, cementoblasts by OCN , PDL fibroblasts by ALP at moderate level, cell clusters/nodules of cementoblasts were observed as the cells approached confluence.Conclusion: Cementoblast-like cells may be cultured in vitro.

关 键 词：大鼠 成牙骨质细胞 细胞培养 生物学特性 鉴定 

分 类 号：Q813.11[生物学—生物工程]