人牙周膜干细胞的初步鉴定及体外成骨诱导

Human Periodontal Ligament Stem Cells Differentiation into Osteoblasts in vitro

目的:体外培养、鉴定人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)并定向诱导分化为成骨细胞,探讨人PDLSCs的多向分化潜能。方法:体外分离、培养人牙周膜细胞,待细胞达一定量后用有限稀释法进行克隆化培养,筛选鉴定牙周膜干细胞(PDLSC),矿化诱导培养21d后检测钙结节形成情况、ALP活性,免疫细胞化学检测骨涎蛋白(BSP)、Ⅰ型胶原表达,RT-PCR检测ALP、BSPmRNA表达。结果:人PDLSCs体外诱导培养21d后可见钙结节形成,成骨细胞相关蛋白及mRNA均阳性表达。结论:人PDLSCs在体外诱导条件下具有成骨潜能。

Objective： To isolate and purify the human periodontal ligament stem cells（PDLSCs） and investigate the differentiation potentials of PDLSC into osteoblasts in vitro. Methods： PDLSCs were isolated and cultivated. PDLSCs of passage 2 was plated at density of 5×10^3/cm^2. At 80% confluence, the medium was changed to an inducing medium. After 21 days of induction, the results were evaluated by Alizarin red S staining, ALP activitv test, immunohistochemistry and RT-PCR. Results： PDLSCs were cultivated and differentiated in vitro with active function, which were manifested by Alizarin red S staining and ALP activity test. Positive expression of collagen Ⅰ , BSP, ALP were confirmed by immunohistochemistry and RT-PCR. Conclusion： PDLSCs can be induced into osteoblasts in vitro. PDLSCs have the capability of multilineage differentiations.