大鼠原代脊髓微血管内皮细胞的体外分离培养及鉴定  

Isolation culture and identification of primary rat spinal cord microvascular endothelial cells in vitro

作　　者：张帆 李柏霖 刘海琴 唐元瑜[2] ZHANG Fan;LI Bolin;LIU Haiqin;TANG Yuanyu(College of Acupuncture and Moxibustion,Fujian University of TCM,Fuzhou 350122;College of Traditional Chinese Medicine,Fujian University of TCM,Fuzhou 350122;Shanghai Changzheng Hospital,the Second Affiliated Hospital to Naval Medical University,Shanghai 200003,China)

机构地区：[1]福建中医药大学针灸学院,福州350122 [2]福建中医药大学中医学院,福州350122 [3]海军军医大学第二附属医院上海长征医院,上海200003

出　　处：《神经解剖学杂志》2023年第5期563-567,共5页Chinese Journal of Neuroanatomy

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

摘　　要：目的:建立一种简单高效的大鼠原代脊髓微血管内皮细胞培养方法。方法:分离大鼠脊髓组织,经剪碎、细胞筛网过滤、Ⅱ型胶原酶消化后接种培养。通过细胞形态学观察、FⅧRag免疫细胞化学染色鉴定所培养的目的细胞。结果:接种的脊髓微血管段呈“短棍样”,培养24 h后有少量短梭形细胞从血管段周围迁移爬出,48 h后逐渐形成“岛屿状”的细胞集落,72 h后细胞铺满皿底,呈典型的单层、“鹅卵石样”、镶嵌式排列生长;细胞FⅧRag免疫细胞化学染色显示细胞质呈棕红色,表达为阳性。结论:该方法可成功分离培养出活性好、纯度高的大鼠原代脊髓微血管内皮细胞。Objective:To establish a simple and efficient primary culture method for rat spinal cord microvascular endothelial cells.Methods:Rat spinal cords were dissected and minced,and the cells were filtered through a sieve and cultured after centrifugation with a bovine serum albumin density gradient and collagenase type II digestion.The target cells were identified by cell morphology observation and immunocytochemical staining of factorⅧrelated antigen(FⅧRag).Results:The cultured spinal cord microvessel segments showed a“beaded”appearance.After 24 hours of culture,a small number of short spindle-shaped cells crawled out from the adherent tissue after 48 hours,and thoroughly covered the bottom of the plate after 72 hours.The cell morphology was full,typical of a single layer,“cobble stone-type”,and embedded wall growth;immunocytochemical staining showed positive expression of FⅧRag.Conclusion:This method can successfully isolate and culture primary rat spinal cord microvascular endothelial cells with high purity and good vitality.

关 键 词：脊髓微血管内皮细胞 原代培养 第Ⅷ因子相关抗原 免疫细胞化学染色 大鼠 

分 类 号：R-33[医药卫生]