摘要
目的:采用差异过筛法培养纯度高、活性好的原代大鼠脑微血管内皮细胞(BMECs)。方法:取4周龄SD大鼠脑皮质,经剪碎、细胞筛网过滤、收集网下滤过物、Ⅱ型胶原酶消化后,置于CO_(2)培养箱中进行原代培养。通过细胞形态学观察、血管性血友病因子(vWF)免疫细胞化学染色鉴定所培养的目的细胞。结果:培养24 h后血管段周围爬出短梭形的细胞;48 h后岛屿状的细胞集落形成;72 h后细胞铺满瓶底,呈典型的单层、铺路石样、镶嵌式贴壁生长;免疫细胞化学染色显示,所培养细胞的细胞质呈现棕红色,vWF表达为阳性。结论:差异过筛法能够成功分离培养出原代大鼠BMECs。
AIM:To establish a method for isolating and culturing primary rat brain microvascular endothelial cells(BMECs)with high purity using the differential sieving technique.METHODS:Brain tissue from 4-week-old SD rats was minced,passed through a cell strainer,and digested with type Ⅱ collagenase to obtain brain microvascular seg-ments,which were then placed in a CO_(2) incubator for primary culture.The cells were identified by morphological observa-tion and immunocytochemical staining for von Willebrand factor(vWF).RESULTS:After 24 h of culture,short spindle-shaped endothelial cells migrated out from the microvascular segments.Forty-eight hours later,the cells formed island-like colonies.After 72 h,the cells covered the bottom of the container and exhibited a typical cobblestone-like pattern.Immunocytochemical staining for vWF showed brownish-red cytoplasm,indicating positive expression.CONCLUSION:The differential sieving method can successfully isolate and culture primary rat BMECs with exceptional purity.
作者
刘海琴
张帆
李柏霖
唐元瑜
LIU Haiqin;ZHANG Fan;Li BoLin;TANG Yuanyu(Department of Respiratory and Critical Care Medicine,Shanghai Changzheng Hospital,Second Affiliated Hospital of Na-val Military Medical University,Shanghai 200003,China;College of Acupuncture and Moxibustion,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;College of Traditional Chinese Medicine,Fujian University of Tra-ditional Chinese Medicine,Fuzhou 350122,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第9期1724-1728,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81072714)
福建省自然科学基金资助项目(No.2017J01545)。
关键词
脑微血管内皮细胞
原代培养
差异过筛法
血管性血友病因子
大鼠
brain microvascular endothelial cells
primary culture
differential sieving method
von Wille-brand factor
rats
