【摘要】 目的建立脐静脉内皮细胞(HUVECs)的体外血管生成的三维培养模型。方法以酶消化法获得以HUVECs为主的混合细胞悬液,经反复贴壁法分离纯化HUVECs,通过免疫细胞化学染色鉴定。采用夹心培养法构建HUVECs的体外血管生成三维培养模型。结果HUVECs在胶原凝胶之间,24 h左右血管样结构的分支之间相互沟通连接形成复杂的网状结构,2 d后凝胶内的细胞开始退化,3 d发现大部分细胞出现崩解。24 h凝胶经H-E染色可见细胞索及原始血管腔形成。结论成功建立体外血管生成的三维培养模型。夹心培养法是较为成熟的体外血管生成的三维培养方法,具有操作简便、三维血管样结构形成数量多、网络结构复杂等特点。更多还原

【Abstract】 Objective To construct a three-dimension(3D) in vitro model for angiogenesis of HUVECs.Methods Single-cell mixing suspension most of which are HUVECs was obtained by filling Collagenase into human umbilical vein.HUVECs were purified by plating cells to the cultivate plating repeatingly.The purified cells were identified by immunocellochemistry.Three-dimensional vascular-like structure was acquired in three-dimensional culture system.Result HUVECs were cultured in the collagen gel,in the first 24 h,the complicated three-dimensional vascular-like structure was emerged,and in the second day,the three-dimensional structure began to degenerate,the third day the three-dimensional structure was disaggregation.In the 24th hour,the cell cable and primitive lumen of blood vessel were confirm by H-E staining through paraffin wax microtome section.Conclusion The three dimension(3D) in vitro model for angiogenesis were constructed successfully.The three-dimensional cell culture system in vitro is a convenient method which has merit in the growth of endothelial cell,the formation of tube-meshworks and the three-dimensional meshwork,and especially suits for the study of the effects of the factors on the angiogenesis.更多还原