摘要
目的 探寻耳蜗微血管内皮细胞的分离与体外培养新方法。方法 选取豚鼠 4只 ,无菌条件下分离其耳蜗血管纹和螺旋韧带 ,37℃下 ,Ⅳ型胶原酶 (0 .5mg/ml)消化 3h ,将消化获得的细胞接种后用内皮细胞条件培养液进行培养 ,反复刮除杂细胞。用ABC免疫酶染色法检测Ⅷ因子相关抗原来鉴定培养的内皮细胞 ,设阳性对照、阴性对照和空白对照。结果 所培养的耳蜗微血管内皮细胞纯度极高 ,胞浆内均含有内皮细胞特有的Ⅷ因子相关抗原。结论 本实验培养获得了纯净的耳蜗微血管内皮细胞 ,其方法简便、可操作性强 ,为血迷路屏障的体外研究奠定了坚实的基础。
Objective To obtain a new approach of dissociation and culture of capillary endothelial cell from the stria vascularis and spiral ligament of guinea pig inner ear.Methods The stria vascularis and spiral ligament segments were got from 4 guinea pigs.Capillaries from the stria vascularis and spiral ligament were treated by repeatedly curettement and culture in the conditioned medium obtained from lung capillary endothelial cell strain.The endothelial origin of the cultured cells was confirmed by immunoenzymatic staining demonstration of the presence of Factor Ⅷ-related antigen.Results More than 99% of the cultured cells,in whose cytoplasm the factor Ⅷ-related antigen present,were from endothelium. Conclusion We obtained the cultured capillary endothelial cell from the stria vascularis and spiral ligament of guinea pig inner ear in the study.It is a convenient and feasible approach of culture of capillary endothelial cell from the stria vascularis and spiral ligament of inner ear.
出处
《重庆医学》
CAS
CSCD
2003年第11期1447-1449,共3页
Chongqing medicine
基金
国家自然科学基金资助项目 (39870 834)
