Separation, culture and identification of SD rat corpus cavernosal endothelial cells
Separation, culture and identification of SD rat corpus cavernosal endothelial cells
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DOI：
10.1111/j.1439-0272.2012.01274.x
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发表时间：
2012-03
期刊：
影响因子：
2.4
通讯作者：
J. Chen;C. L. Sun;Z. Chen;H. Xiao;T. Qi;X. Li;X. Tao;B. Zhang
中科院分区：
文献类型：
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作者：
J. Chen;C. L. Sun;Z. Chen;H. Xiao;T. Qi;X. Li;X. Tao;B. Zhang
本研究旨在探讨SD大鼠海绵体血管内皮细胞的分离、培养和鉴定方法。海绵体组织取自雄性SD大鼠。采用酶消化法分离CCECs。免疫磁珠和流式细胞仪细胞分选获得纯化的细胞，在EMG-2培养液中传代。四唑盐3‐[4，5‐dimethylthiazol‐2‐yl]‐2，5‐diphenyltetrazolium溴化法测定细胞生长曲线。用von Willebrand因子(VWF)进行免疫荧光鉴定，并用流式细胞仪检测vWF表达的阳性百分率。显微镜下可见单形鹅卵石样细胞。免疫磁珠可获得较高的纯度。培养2天后，细胞进入对数生长期，第5天达到平台期。VWF在胞浆中呈阳性表达。流式细胞仪检测细胞纯度为95.8%。采用酶消化法、免疫磁珠法和流式细胞仪细胞分选法，成功分离培养SD大鼠脐静脉内皮细胞。
The aim of the study is to investigate the methods of separation, culture and identification of Sprague Dawley (SD) rat corpus cavernosal vascular endothelial cells (CCECs). Cavernosal tissues were isolated from male SD rats. Enzymatic digestion was applied to separate CCECs. Purified cells were obtained using immunomagnetic beads and flow cytometric cell sorting and subcultured in EMG‐2 medium. The growth curve of CCECs was measured by the tetrazolium salt 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide assay. The cells were identified by von Willebrand factor (vWF) using immunofluorescence, and the positive percentage of vWF expression was detected by flow cytometry. The monomorphic cobblestone‐like cells were observed by microscopy. High purification was obtained using immunomagnetic beads. After 2 days of incubation, cells entered the logarithmic growth phase and reached a plateau on the fifth day. The vWF expression in cytoplasm was positive. The purity of cells was 95.8%, which was tested by flow cytometry. SD rat CCECs can be separated and cultured successfully by the method of enzymatic digestion, immunomagnetic beads and flow cytometric cell sorting.