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目的 :比较体外成骨诱导培养的小鼠胚胎10.5 d(E10.5)的上颌突间充质细胞与体内发育的上颌突间充质细胞的成骨分化差异。方法:体外培养E10.5和E17.5的小鼠上颌突间充质细胞,观察其细胞形态。取体外培养3 d的E10.5原代细胞进行成骨诱导培养7 d,然后应用免疫荧光、qPCR等方法比较其与体外培养3 d的E17.5原代细胞的成骨分化差异。采用SPSS 20.0软件包对数据进行成组样本t检验。结果:E10.5和E17.5的小鼠上颌突细胞在体外呈贴壁生长,细胞呈多边形、椭圆形。E17.5的小鼠上颌突原代间充质细胞在体外培养时,增殖速度较E10.5的小鼠上颌突细胞快。E10.5的小鼠上颌突间充质细胞成骨诱导7 d后,成骨标志物Runx2和OCN的蛋白表达与未成骨诱导的E17.5上颌突间充质细胞表达相似,成骨标志物Runx2、OCN和OPN的mRNA表达和未成骨诱导的E17.5上颌突细胞表达相似。成骨诱导培养14 d的E10.5的小鼠上颌突细胞与成骨诱导7 d的E17.5上颌突细胞形成的钙结节无显著差异。结论:E10.5的小鼠上颌突间充质细胞体外成骨诱导培养,能较好模拟上颌突细胞体内成骨发育过程,为研究颌骨发育提供了合适的细胞模型。
OBJECTIVE: To compare the osteogenic differentiation between maxillary mesenchymal cells and maxillary mesenchymal cells cultured in vitro induced by osteogenic induction of mouse embryos 10.5 d (E10.5). Methods: The mouse maxillary protrusion mesenchymal cells of E10.5 and E17.5 were cultured in vitro and their cell morphology was observed. The primary cultured E10.5 cells cultured in vitro for 3 days were cultured for 7 days. Then, the osteogenic differentiation of primary E17.5 cells cultured in vitro for 3 days were compared by immunofluorescence and qPCR. Data was subjected to group t-test using SPSS 20.0 software package. Results: The mouse maxillary cells of E10.5 and E17.5 showed adherent growth in vitro with polygonal and oval cells. E17.5 mouse maxillary process of primary mesenchymal cells cultured in vitro, the proliferation rate of E10.5 mouse maxillary cells faster. After osteoblast induction of mouse maxillofacial mesenchymal cells in E10.5 for seven days, the expressions of Runx2 and OCN were similar to those of non-osteoinductive E17.5 maxillary mesenchymal cells. Osteoblast markers The mRNA expression of Runx2, OCN and OPN was similar to that of non-osteoinduced E17.5 maxillary cells. There was no significant difference in calcium nodules formed between osteoblast-induced E10.5 mouse maxillary cells cultured for 14 days and E17.5 maxillary cells cultured for 7 days. CONCLUSION: E10.5 mouse maxillary mesenchymal cells cultured in vitro can well simulate the process of osteoblast development in vivo and provide a suitable cell model for the development of maxilla.