血管平滑肌细胞的原代培养采用贴块法进行。选用SD大鼠(南方医科大学南方医院实验动物中心提供)断头处死, 取胸主动脉中膜以贴块法培养于含10% 胎牛血清的DMEM培养基中, 置于37℃、5% CO2孵箱中培养, 一周后可见细胞从组织块边缘爬出, 2～3周出现致密细胞层, 此时即可传代, 传代细胞呈典型的“峰与谷”样生长, 细胞经抗α- actin抗体鉴定为平滑肌细胞。实验所用的血管平滑肌细胞均为第4代传代细胞深圳英语翻译。
The primary culture of vascular smooth muscle cell used attachment-block method. SD rats (supplied by South Medical University South Hospital Experimental Animal Center) were executed by decollation, took thoracic aorta mesolamella and cultured by attachment-block in DMEM culture media with 10%fetal bovine serum, then put into 37℃, 5% CO2 incubator. One week later, cells can be seen crawling from the edge. Dense cell layer can be formed after 2-3 weeks which can passage. The passage cells were in typical “apex and vale” growth, and were identified by anti α- actin antibody as smooth muscle. Vascular smooth muscle cells used in lab were all the 4th generation of passage cells.
VSMCs在DMEM培养基中培养汇满至80％, 用含有0.2％小牛血清DMEM培养基培养24h, 使细胞处于同步状态, 更换实验用培养基培养; 60min后消化细胞, 使用PI标记, 资料用CELLTIL细胞周期分析软件处理英语翻译。
when the VSMCs in DMEM culture media filled 80％, cultured them with 0.2％ Bovine Serum DMEM culture media for 24h, which let the cells in the same status, and then used the culture media for experiment; Digested for 60min, and then marked by PI. The data were dealt with CELLTIL cell cycle analysis software.
