摘要
目的:体外分离纯化人牙周膜干细胞,研究其体外矿化能力,为牙周组织工程提供可靠的种子细胞来源。方法:采用有限稀释法克隆分离纯化人牙周膜干细胞,将获得的第5代细胞用矿化液连续培养,观察矿化情况,进行矿化结节Von kossa染色;通过免疫组织化学染色技术检测诱导后细胞骨黏连素(ON)、骨桥蛋白(BSP)、Ⅰ型胶原(COLⅠ)、Ⅲ型胶原(COLⅢ)的表达。结果:人牙周膜细胞在体外可以叠层生长,形成肉眼可见的灰白色结节,Von kossa染色显示结节内有钙盐沉积,该细胞诱导21 d后有矿化相关蛋白COLⅠ、COLⅢ,BSP、ON的阳性表达。结论:人牙周膜干细胞在体外矿化液作用下出现向成骨细胞的分化。
AIM：To isolate and purify the periodontal ligament stem cells from human third molars and investigate inducing differentiation of PDLSCs in mineralized medium. METHODS ： PDLSCs were isolated by limited dilution of culture cells for single cell clone. PDLSCs were induced with minerralized conditional medium for 21 days. Differentiation of PDLSCs were assessed. Mineralized potential was studied by Von - Kossa staining . COL Ⅰ,COLⅢ, BSP ,ON were examined by immunohistochemistry stains. RESULTS： PDLSCs formed in white calcified nodules by induced with minerralized conditional medium. The calcified nodules were stained by Von - Kossa. COLⅠ, COL Ⅲ, BSP,ON were positive stronger in the induces cells. CONCLUSION： PDLSCs could be induced to differentiate into osteoblasts in minerralized conditional medium in vitro culture.
出处
《牙体牙髓牙周病学杂志》
CAS
2007年第4期195-199,共5页
Chinese Journal of Conservative Dentistry
基金
陕西省自然科学研究计划(2003C2042)
关键词
人牙周膜干细胞
细胞克隆
诱导分化
免疫细胞化学
periodontal ligament stem cell
cell cloning
inducing differentiation
immunohistochemistry
