Shen Yuan-yuan, Chen Ke, Xu Nuo
Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jan;30(1):96-9.
To test the capacity of the stem cells derived from human exfoliated deciduous teeth in in vitro differentiation into osteoblasts.
Stem cells were isolated from the exfoliated deciduous teeth of healthy children and sorted into CD34(+)/CD117(+) cells and the remaining mixed cells using flow cytometry. After in vitro cell culture, the differentiation capacity into osteoblasts of the two groups of cells was evaluated by detecting the markers of osteoblasts using immunocytochemical techniques and fluorescent quantitative PCR. Mineralization assay was performed to identify the cell differentiation.
The cells isolated by typsin digestion grew in the manner of fibroblasts. After a 30-day culture of the two groups of cells, immunocytochemistry detected the expressions of osteoblast markers RUNX-2, OC, and BSP. After 40 days of cell culture, the mRNA expressions of RUNX-2, OC and BSP genes were significantly different between the two groups. At day 50 of cell culture, the CD34(+)/CD117(+) cells exhibited positivity for von Kossa's staining and alizarin red staining, but the mixed cells showed negative staining results.
The purified CD34(+)/CD117(+) stem cells derived from exfoliated deciduous teeth of healthy children possess the capacity to differentiate into osteoblasts and form calcium deposits and mineralized nodules in vitro.
检测人脱落乳牙来源的干细胞在体外分化为成骨细胞的能力。
从健康儿童的脱落乳牙中分离干细胞，采用流式细胞术将其分为CD34(+)/CD117(+)细胞和其余混合细胞。体外细胞培养后，通过免疫细胞化学技术和荧光定量PCR检测成骨细胞标志物，评估两组细胞向成骨细胞的分化能力。进行矿化试验以鉴定细胞分化情况。
经胰蛋白酶消化分离的细胞呈成纤维细胞样生长。两组细胞培养30天后，免疫细胞化学检测到成骨细胞标志物RUNX-2、OC和BSP的表达。细胞培养40天后，两组之间RUNX-2、OC和BSP基因的mRNA表达存在显著差异。细胞培养50天时，CD34(+)/CD117(+)细胞对冯科萨染色和茜素红染色呈阳性反应，但混合细胞染色结果为阴性。
从健康儿童脱落乳牙中纯化得到的CD34(+)/CD117(+)干细胞具有在体外分化为成骨细胞并形成钙沉积和矿化结节的能力。
