小鼠上颌突间充质细胞体内成骨发育和体外诱导成骨分化的比较

doi:10.19439/j.sjos.2017.03.009

上海口腔医学 ›› 2017, Vol. 26 ›› Issue (3): 277-280.doi: 10.19439/j.sjos.2017.03.009

小鼠上颌突间充质细胞体内成骨发育和体外诱导成骨分化的比较

郑军, 王雷, 母东亮, 杜玉芳, 刘丽佳, 郑杰

大庆龙南医院·齐齐哈尔医学院第五附属医院口腔外科,黑龙江大庆 163411

收稿日期:2016-09-12 修回日期:2016-11-21 出版日期:2017-06-25 发布日期:2017-07-05
通讯作者: 郑军,E-mail： 70806744@qq.com
作者简介:郑军(1982-),硕士研究生,主治医师

Osteogenic development in vivo and osteogenic differentiation in vitro of mouse maxillary primordium mesenchymal cells: a comparative study

ZHENG Jun, WANG Lei, MU Dong-liang, DU Yu-fang, LIU Li-jia, ZHENG Jie

Department of Oral Surgery, Daqing General Hospital Group Longnan Hospital. Daqing 163411, Heilongjiang Province, China

Received:2016-09-12 Revised:2016-11-21 Online:2017-06-25 Published:2017-07-05

摘要/Abstract

摘要： 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。

关键词: 上颌突, 间充质细胞, 体内发育, 体外成骨分化

Abstract: PURPOSE: To compare the differences between the osteogenic development in vivo and osteogenic differentiation in vitro of mouse maxillary primordium mesenchymal cells (MPMCs). METHODS: E10.5 and E17.5 mouse MPMCs were cultured in vitro to observe cell morphology. E10.5 primary MPMCs, after culturing in vitro for 3 days, were cultured in osteogenic differentiation in vitro for another 7 days. Then immunofluorescence and qPCR were used to compare the difference of osteogenic differentiation with E17.5 primary MPMCs cultured in vitro for 3 days. SPSS 20.0 software package was used for independent samples t test. RESULTS: E10.5 and E17.5 mouse MPMCs adhered to dish when cultured in vitro, and the cells exhibited polygonal or oval shape. The proliferation of E17.5 mouse MPMCs was faster than that of E10.5 MPMCs. After 7 days of osteogenic induction, the expression of Runx2 and OCN proteins, two osteogenic markers, in E10.5 mouse MPMCs was similar to E17.5 cells without osteogenic induction. The mRNA expression of Runx2, OCN and OPN also showed similar expression patterns, and there was no significant difference in the calcium nodules formation between E10.5 MPMCs and E17.5 MPMCs. CONCLUSIONS: In vitro osteogenic induction of E10.5 mouse MPMCs can mimic osteogenic development process of MPMCs in vivo, and provide a suitable cell model for the study of jaw development.

Key words: Maxillary primordium, Mesenchymal cells, Development in vivo, Osteogenic differentiation in vitro

中图分类号:

R739.8

郑军, 王雷, 母东亮, 杜玉芳, 刘丽佳, 郑杰. 小鼠上颌突间充质细胞体内成骨发育和体外诱导成骨分化的比较[J]. 上海口腔医学, 2017, 26(3): 277-280.

ZHENG Jun, WANG Lei, MU Dong-liang, DU Yu-fang, LIU Li-jia, ZHENG Jie. Osteogenic development in vivo and osteogenic differentiation in vitro of mouse maxillary primordium mesenchymal cells: a comparative study[J]. Shanghai Journal of Stomatology, 2017, 26(3): 277-280.

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小鼠上颌突间充质细胞体内成骨发育和体外诱导成骨分化的比较

Osteogenic development in vivo and osteogenic differentiation in vitro of mouse maxillary primordium mesenchymal cells: a comparative study

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