摘要
目的研究体外培养的人牙周膜细胞群（hPDLP）向成骨和成脂方向分化的潜能,为牙周组织工程提供可靠的种子细胞来源。方法组织块法分离培养人牙周膜细胞群,流式细胞术检测间充质干细胞标记CD146和STRO-1的表达;利用茜素红、油红O染色、免疫组化以及反转录聚合酶链反应（RT-PCR）等检测hPDLP的多向分化标志。结果第1代hPDLP的CD146和STRO-1阳性率分别是27.20%±3.98%和4.23%±4.08%;经矿化诱导可以形成矿化结节,有钙盐沉积;经成脂诱导可见特异性脂滴形成,特异性转录因子过氧化物酶体激活物增生受体2（PPARγ2）和脂蛋白脂酶（LPL）表达上调。第8代hPDLP相对第1代的矿化能力没有差异,但成脂方向分化潜能减弱。结论体外培养的hPDLP具有向成骨和成脂样细胞分化潜能,第1~3代细胞群明显具有牙周膜干细胞的多向分化潜能优势。
Objective To explore the multi-differentiated capability of human periodontal ligament cell population （hPDLP）, and provide a theoretical basis for the periodontal regeneration by tissue engineering technique. Methods hPDLP was cultured from periodontium of human tooth by the outgrowth method. STRO-1 and CD146 expression were investigated by flow cytometry. hPDLP was induced to odontogenic/osteogenic-like and adipogenic-like cell. The multilineage differentiation capacities of hPDLP were evaluated by alizarin red stain, oil red O stain, anti-CD146 and STRO-1 immunocytochemistry, and reverse-transcriptase polymerase chain reaction analysis. Results hPDLP was isolated from human periodontium and most of the cells retained their fibroblastic spindle shape. hPDLP can be induced into osteoblast-like cells and adipocyte-like cells, and calcium deposition and lipid droplets were detected perspectively. And the eighth generation of hPDLP had weaker potential into adipocyte-like cells than the first passage, however, there was no difference to the aspect of calcification ability between the two passages. Conclusion hPDLP cultured in vitro can differentiate into adipocytes and osteoblasts, and the first to third passage cells may have the predominance of differentiation potential.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2010年第2期185-189,共5页
West China Journal of Stomatology
基金
广东省科技计划项目重大科技专项基金资助项目(2006B35801010)
关键词
人牙周膜细胞群
分化潜能
矿化
诱导
human periodontal ligament cell population
differentiation potential
calcification
induction
