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作 者:洪海霞[1] 张彦明[1] 孙裴[1] 苏正元[1] 魏中锋[1] 张永国[1]
机构地区:[1]西北农林科技大学动物科技学院,陕西杨凌712100
出 处:《西北农林科技大学学报(自然科学版)》2004年第5期1-4,共4页Journal of Northwest A&F University(Natural Science Edition)
基 金:国家自然科学基金项目(30270988)
摘 要: 采用1g/L胶原酶灌注猪脐静脉,消化分离内皮细胞并在适宜条件下培养。对培养细胞的生长特性进行检测,并对细胞进行第 因子相关抗原(F- RAg)和扫描电镜形态学鉴定。结果表明,70%细胞在24h内贴壁,第3~5天生长迅速,第4~5天长成单层。培养的细胞呈单层贴壁生长,典型铺路石状,第 因子相关抗原(F- RAg)检测阳性,证明培养的细胞为血管内皮细胞。Swine umbilical cord was sterilized by 75% ethyl alcohol about 1 min and umbilicus veins were washed by PBS.1 g/L callagenase Ⅰ was poured into umbilicus vein and digested 5 min at 37 ℃.Endothelial cells obtained was collected by centrifugation,cultured in M199+20% fetal calf serum.The plating efficiency to plastic was 70%-75% for 24 hours in callagenase dispersed cells.A confluent monolayer was former by 4-5 days.The the endothelial cells looked like cobblestones and were stained positively for factor Ⅷ relative antigen,which is feature of endothelial cells.It shows cells cultured are umbilicus veins cells.
关 键 词:脐静脉 血管内皮细胞 分离 细胞培养 胶原酶 猪瘟病毒 第Ⅶ因子相关抗原 发病机理
分 类 号:S858.28[农业科学—临床兽医学]
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