改良组织块培养法提取原代大鼠脑微血管内皮细胞及鉴定  

Extraction and identification of primary rat brain microvascular endothelial cells by improved tissue block culture method

作　　者：张帆[1] 李柏霖 池茗 刘海琴 唐元瑜[2] Zhang Fan;Li Bolin;Chi Ming;Liu Haiqin;Tang Yuanyu(Dept of Acupuncture and Moxibustion,College of Acupuncture and Moxibustion,College of Integrated Traditional Chinese and Western Medicine,Fujian University of TCM,Fuzhou 350122;Basic Theory of TCM Discipline,College of TCM,College of Integrated Traditional Chinese and Western Medicine,Fujian University of TCM,Fuzhou 350122;Department of Clinical Medicine,College of Integrated Traditional Chinese and Western Medicine,Fujian University of TCM,Fuzhou 350122;Dept of Respiratory and Critical Care Medicine,Shanghai Changzheng Hospital Affiliated Naval Military Medical University,Shanghai 200003)

机构地区：[1]福建中医药大学针灸推拿学院针推系,福州350122 [2]福建中医药大学中医学院中医基础理论学科,福州350122 [3]福建中医药大学中西医结合学院临床医学系,福州350122 [4]海军军医大学上海长征医院呼吸与危重症医学科,上海200003

出　　处：《安徽医科大学学报》2025年第1期10-14,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81072714);福建省自然科学基金(编号:2017J01545)。

摘　　要：目的探究脑微血管组织块培养法提取原代大鼠脑微血管内皮细胞并对其进行鉴定。方法取4周龄SD大鼠脑皮层,经过筛网、预消化、微血管组织块固化处理后,置于CO_(2)培养箱中进行原代培养。通过细胞形态学观察、第Ⅷ因子相关抗原免疫细胞化学染色法鉴定所培养的目的细胞。结果体外培养48 h后短梭形细胞从脑微血管段周围爬出;72 h后岛屿状的细胞团簇形成;96 h后团簇融合,细胞呈典型的单层铺路石样镶嵌式排列生长。第Ⅷ因子相关抗原免疫细胞化学染色法检测显示,细胞胞质呈棕红色,表达为阳性,细胞核被苏木精衬染成蓝黑色。结论脑微血管组织块培养法能够成功提取原代大鼠脑微血管内皮细胞。Objective To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect.Methods Brain tissue from 4-week-old Sprague Dawley rats was screened,pre-digested and solidified to obtain brain microvascular segments.These segments were subsequently placed in a CO_(2)incubator for primary culture.The target cells were identified by cell morphology and immunocytochemical staining for factorⅧ-related antigen.Results After a 48-hour culture period in vitro,the short spindle cells crawled out from around the brain microvascular segments.After 72 hours,island-like cell culsters formed.After 96 hours the clusters fused and the cells formed a typical monolayer,cobble stone-like,and mosaic arrangement.FactorⅧ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red,indicating positive expression;DAB stained the nucleus,showing blue-dark.Conclusion The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.

关 键 词：脑 微血管内皮细胞 原代培养 脑微血管组织块培养法 第Ⅷ因子相关抗原 大鼠 鉴定 

分 类 号：R331.3[医药卫生—人体生理学]