人脐静脉内皮细胞原代培养及鉴定
发布时间：2019-01-03 12:09
【摘要】：目的建立一个详细、易于重复的人脐静脉内皮细胞原代培养操作流程。方法采用胶原酶消化法分离人脐静脉内皮细胞;用倒置显微镜观察细胞形态,免疫荧光法检测内皮细胞标志分子,Western blotting检测内皮细胞标志酶——一氧化氮合酶的含量。结果 0.2%胶原酶Ⅰ在37℃条件下消化5~13 min可得到高纯度(95%)人脐静脉内皮细胞;在倒置显微镜下,人脐静脉内皮细胞呈典型的鹅卵石样,细胞核明显;Ⅷ因子和CD31的免疫荧光检测呈阳性;Western blotting结果显示,分离到的人脐静脉内皮细胞中有一氧化氮合酶持续合成。结论建立的人脐静脉内皮细胞原代培养和鉴定的操作流程简单易学,且在形态、标志分子和关键性标志酶3个方面提供细胞鉴定的方法和参考数据。
[Abstract]:Objective to establish a detailed and reproducible primary culture procedure for human umbilical vein endothelial cells (HUVECs). Methods Human umbilical vein endothelial cells were isolated by collagenase digestion, the morphology of endothelial cells was observed by inverted microscope, and the content of nitric oxide synthase (NOS), a marker of endothelial cells, was detected by immunofluorescence assay. Results 0.2% collagenase I could obtain high purity (95%) of human umbilical vein endothelial cells after digesting at 37 鈩，
本文编号：2399342
[Abstract]:Objective to establish a detailed and reproducible primary culture procedure for human umbilical vein endothelial cells (HUVECs). Methods Human umbilical vein endothelial cells were isolated by collagenase digestion, the morphology of endothelial cells was observed by inverted microscope, and the content of nitric oxide synthase (NOS), a marker of endothelial cells, was detected by immunofluorescence assay. Results 0.2% collagenase I could obtain high purity (95%) of human umbilical vein endothelial cells after digesting at 37 鈩，
本文编号：2399342
本文链接：https://www.wllwen.com/xiyixuelunwen/2399342.html
最近更新
教材专著
