摘要
目的建立一种改良的血管内皮细胞培养方案,为体外培养血管内皮细胞提供稳定而有效的实验方法。方法在无菌条件下,应用输液延长管置入脐带并注入2/3容积的0.1%Ⅱ型胶原酶消化、分离人脐静脉内皮细胞。用倒置相差显微镜观察细胞生长,免疫细胞化学法鉴定培养细胞。结果原代培养的细胞30 min开始贴壁生长,24￣48 h生长最快,3￣5 d呈铺路石样排列。免疫细胞化学可见培养细胞胞浆中人Ⅷ因子相关抗原呈阳性反应,证实培养的细胞为血管内皮细胞,且纯度达97.5%。结论用本实验中的方法消化、分离血管内皮细胞可获得高纯度的内皮细胞,细胞数量多、污染少且成活率高。
【Objective】To develop a method of culturing vascular endothelial cells（） from human umbilical veins,to provide a stable and effective experimental method for culturing vascular endothelial cells in vitro.【Methods】The interior of the umbilical vein was digested by infusion of 2/3 volume 0.1 % typeⅡ collagenase.VECs were isolated in aseptic condition.Then the growth of cells was observed under an inverted phase contrast microscope,the cultured cells were identified by immunocytochemistry stain.【Results】The primary cultured cells began to grow attaching to wall about 30 minutes after inoculating and arrayed like pitching stone in 3-5days under an inverted phase contrast microscope.Ⅷ factor in the VECs showed positive reaction by immunocytochemistry,the purity of primary generation VECs assessed by immune staining was 97.5 %,which were identified as VECs.【Conclusion】 By our experimental method,high yield of pure VECs can be harvested with high survival rate.This method provides effective experimental model for VECs research.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第6期848-850,共3页
China Journal of Modern Medicine
关键词
脐静脉内皮细胞
改良培养
免疫细胞化学
倒置相差显微镜
umbilical vein endothelial cells
improved culture
immunocytochemistry
inverted phase contrast microscope
