C57BL/6J小鼠胚胎腭突体外培养模型的建立  

Establishment of palatal organ culture model of C57BL/6J mouse embryos in vitro

作　　者：张岱尊[1] 庄翠竹[1] 肖文林[1] 许尧祥[1] 王双义[1] 

机构地区：[1]青岛大学医学院附属医院口腔科,山东青岛266003

出　　处：《上海口腔医学》2013年第2期132-136,共5页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(81070817);山东省自然科学基金(ZR2010HM054);青岛市科技计划资助项目(08-2-1-5-nsh)~~

摘　　要：目的:建立近交系C57BL/6J小鼠胚胎腭突体外培养模型,为研究胚胎腭突发育提供条件。方法:无菌条件下,在体视显微镜下取怀孕13.5 d(GD13.5)的胚胎腭突,进行培养。分别于培养6、24、48 h后取出固定,行组织切片苏木精-伊红(HE)染色和扫描电镜(SEM)观察,每个时间点分别12对。结果:细胞核染色显示,6 h时,两侧腭突未融合;24 h时,两侧腭突开始融合,但是中间仍有部分腭中嵴上皮(MEE)细胞残留;48 h时,两侧腭突完全融合,中间的MEE完全消失。扫描电镜显示,6 h时,两侧腭突之间有一定裂隙;24 h时,两侧腭突已接触但能观察到腭中缝(MES);48 h时,两侧腭突已完全融合,MES消失。结论:本方法为研究腭裂发病机制提供了一个良好的体外模型。PURPOSE： To establish palatal organ culture model of C57BL/6J mouse embryos in vitro and provide platform for study of embryo palatal development. METHODS： The mouse palatal shelves were harvested under sterilization from a female mouse of gestation day （GD） 13.5 by stereoscopic microscope and cultured in vitro. Totally 36 pairs of palatal shelves were divided into three groups equally and cultured 6 h, 24 h and 48 h, respectively. Finally, all palatal shelves were embedded and stained by Hematoxylin and Eosin （HE） and subjected to scanning electron microscope （SEM） analysis. RESULTS： The results of HE dyeing showed that the palatal shelves did not fuse on 6 h group, and began to fuse on 24 h group, but still had some medial edge epithelial （MEE） cells remained. The palatal shelves completely fused and all the MEE cells disappeared on 48 h group. The results of SEM showed that there was a gap between palatal shelves on 6 h group. The palatal shelves began to contact and form the medial epithelial seam （MES） on 12 h group. Finally, palatal shelves completely fused and MES disappeared on 48 h group. CONCLUSION： This method provides an effective way for investigating the etiology of cleft palate in vitro. Supported by National Natural Science Foundation of China （81070817）, Natural Science Foundation of Shandong Province （ZR2010HM054） and Science and Technology Planning Project of Qingdao City（08-2-1-5-nsh）.

关 键 词：C57BL 6J小鼠 腭突培养 扫描电镜 融合 

分 类 号：R782.2[医药卫生—口腔医学]