小鼠胚胎腭突间充质细胞体外培养及生物学特性研究

Biological characteristics of mouse embryonic palatal mesenchymal cells in vitro

目的 研究小鼠胚胎腭突间充质 (EPM)细胞的生物学特性。方法 在显微镜下解剖妊娠第 13天的母鼠胚胎腭突 ,用 0 .2 5 %胰蛋白酶进行消化获得游离分散的EPM细胞 ,在含 10 %胎牛血清的DMEM培养基中进行培养。采用免疫组化方法进行细胞鉴定 ,通过相差显微镜 ,生长曲线及透射显微镜观察细胞形态 ,增殖能力及超微结构。结果 EPM细胞为成纤维细胞样细胞 ,排列无序 ,细胞核大 ,核分裂象多 ,核膜内陷 ,核呈分叶状 ,胞浆含大量线粒体及粗面内质网。免疫组化显示角蛋白标记为阴性 ,S -10 0蛋白及波形蛋白标记为阳性。EPM细胞呈指数生长 ,有较强的增殖能力。结论 EPM细胞体外培养生长及增殖良好 ,可作为腭裂基础研究较理想的研究对象。

Objective To investigate the biological characteristics of mouse embryonic palatal mesenchymal (EPM) cells in vitro.Methods The mouse EPM cells were harvested from a female of day 13 of gestation by microsurgical dissection and digestion with 0.25 % trypsin. The isolated cells were cultured in DMEM medium containing 10% fetal bovine serum with 5% CO 2 and 95% air and identified by immunohistochemical method. The cells morphology, proliferation and ultrastructure were observed .Results Mouse EPM cells resembled fibroblast-like cells with no noticeable organization. Transmission electron microscope observation indicated that the cells contained numerous mitochondria and rough endoplasmic reticulum. Mouse EPM cells were confirmed by the positive result of immunohistochemical examination for Vimentin and S-100 and negative result for Pancytokeratin. The proliferation capability of mouse EPM cells was strong. Conclusion In the culture system of mouse EPM cells established here, the cells exhibited good adherence and proliferation capability, which can regard as ideal experimental cells in basic research of cleft palate.