小鼠血管内皮细胞的原代培养

目的:建立简单有效地获取小鼠血管内皮细胞(VEC)的培养方法。方法:从实验动物、培养基、植块方法、胎牛血清浓度等方面对植块培养法进行改进,应用C57BL小鼠腹主动脉血管组织进行血管内皮细胞的原代培养并传代培养培养小鼠血管内皮细胞,并进行倒置显微镜下观察和免疫组织化学进行鉴定。结果与结论倒置显微镜观察小鼠血管内皮细胞生长情况,培养48h后,可见大部分贴壁细胞连接形成网状,细胞密度较低;3～5d后,内皮细胞岛出现;约7～9d后细胞融合成片,细胞呈单层镶嵌状排列,呈现"鹅卵石样"或"铺路石样",出现接触抑制现象。兔抗鼠抗Ⅷ抗体和荧光素标记抗体鉴定结果显示清晰的血管内皮细胞核。

Culture of Mus Vascular Endothelial Cell in Rats

Objective:To develop a simple and rep roductable method for the isolation and primary culture of mus vascular endothelial cell.Method:The tissue block pasted culture method was imp roved in experiment animal,culture-medium,tissue block method,fetal bovine serum(FBS) and spreading slide method.Mus vascular endothelial cell were primary cultured with lung tissue block pasted methods using lung tissue of C57BL Rats,observe mus vascular endothelial cell with inverted microscope and confirmed by immunohistochemistry.Result&Conclusion:we observed mus vascular endothelial cell.with inverted microscope: adherent cell conjuncted with reticulation when cell cultivation for 48h;endotheliocyte island appeared when cell cultivation for 3-5day;cell conjugated with flakiness and cell rank as monolayer inlay、cobblestone or slabstone,which called as contact inhibition.The result of confirmation of mus vascular endothelial cell is that vascular endothelial cell nucleolus clearly appeared with the help of rabbit antimouse antibody.