猪脐静脉血管内皮细胞的分离与培养

Segregation and culture of endothelial cells from swine umbilicus veins

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.