颌骨骨髓基质细胞与牙周膜细胞的生物学特性比较

目的:比较颌骨骨髓基质细胞与牙周膜细胞的基本生物学特性。方法:分离颌骨来源骨髓基质细胞和牙周膜细胞,进行改良体外原代培养。倒置显微镜观察细胞生长及克隆形成情况;CCK-8检测细胞生长曲线;免疫荧光染色检测STR0-1表达;成脂诱导后检测脂滴形成,矿化诱导后检测碱性磷酸酶的变化并用RT-PCR检测7、14 d成骨相关基因OCN的表达水平。结果:两种细胞体外培养均呈成纤维样细胞外形,能克隆生长,均具有活跃的增殖能力;两种细胞STR0-1表达阳性;成脂诱导后可见脂滴形成;矿化诱导后骨髓基质细胞的碱性磷酸酶活性较强,而且OCN基因表达较早且较强。结论:颌骨来源骨髓基质细胞具有较强的增殖及成骨分化能力,具有干细胞特性,可能是有较大临床应用潜力的组织工程种子细胞。

A comparative study on the biological characteristics of human periodontal ligament cells and orofacial bone marrow stromal cells

AIM: To compare the biological characteristics of human orofacial bone marrow stromal cells and periodontal ligament cells in vitro.METHODS:Primary human orofacial bone marrow stromal cells and periodontal ligament cells(PDLCs) were isolated and cultured in vitro.Cell morphology,growth curve,colony formation ratio(CFR),expression of STRO-1 were analyzed by inverted contrast microscope,cck-8 and immunofluorescence staining.After osteogenic induction,alkaline phosphatase activity and the expression of osteocalcin(OCN) were tested by ELISA and RT-PCR.The adipogenic differentiation potential was identified by oil red O staining.RESULTS: Both BMSCs and PDLCs showed fibroblast-like morphology and clonal growth pattern.They proliferated and expanded rapidly in vitro.Both cells expressed the stem cell marker STRO-1.Alkaline phosphatase activity of BMSCs was higher than that of PDLCs on 7 and 14 days after osteogenic induction.BMSCs expressed OCN gene after 7 days of induction,while PDLCs didn’t express it.Lipid droplets were observed in both cells after adipogenic induction.CONCLUSION: The orofacial BMSCs show stem cell characteristics and have strong proliferative capability and osteogenic differentiation potential,hopefully for clinical application as tissue engineering seeding cell.