摘要
目的 寻找来源方便的内皮细胞 ,解决人工血管内皮化问题。方法 收集胎儿脐血 ,Ficoll离心获得单核细胞 ,置内皮细胞培养液中 ,诱导内皮细胞祖细胞转化为内皮细胞并扩增 ;作光镜、扫描电镜形态学观察 ,免疫细胞化学定性、流式细胞仪定量测定 ;采用旋转贴壁培养法 ,使内皮细胞衬里人工血管腔面。结果 1.脐血内皮细胞祖细胞在培养第 5天 ,细胞扩增 3 6± 1 7倍 ,并开始转化为内皮细胞 ,在以后的二周内保持增殖。 2 .免疫细胞化学检测显示 :内皮细胞培养早期 ,Ⅷ因子、CD34、CD31、VEGFR 2表达均为阳性 ,后期除Ⅷ因子有较高表达外 ,其他指标表达阴性。 3 .形态学观察 :在普通玻璃培养瓶中 ,内皮细胞呈铺路石形、单层贴壁生长 ;应用旋转贴壁培养法 ,内皮细胞呈片状生长 ,并贴壁于人工血管腔面 ,符合人工血管内皮化要求。结论 循环内皮细胞能在体外诱导、培养、扩增 ,并能保持一定的生物学活性 ,可作为人工血管内皮研究 ,有望应用于临床。
Objective To discuss the endothelialization of artificial vessel with easily available endothelial cells.Methods Umbilical cord blood monocytes were isolated by centrifugation,cells were propagated in EC growth media,and developing cultures were observed by optical and scanning electron microscopy.Endothelial cells were immunostained for qualitative assay and flow cytometry studied for quantitative assay.ECs were lined to PTFE prosthesis by rotation adherent culture.Results The progenitors were proliferated by up to 3 6±1 7 times,which transduced to endothelial cells on 5th day during culture,and kept proliferation within 2 weeks.Immunostaining of EC was positive for CD31,CD34,factor Ⅷ and VEFGR 2 at premature in culture,which was negative for all but factor Ⅷ at later culture.Morphological observation:EC had the typical cobblestone appearance,and monolayer culture,ECs were lined to PTFE prostheses by rotation adherent culture.Conclusion We conclude the circulating endothelial cells can be induced and propagated during culture in vitro,and studied for endothelialization of artificial vessel in vivo.
出处
《江苏医药》
CAS
CSCD
北大核心
2002年第7期492-493,F002,共3页
Jiangsu Medical Journal
基金
上海市卫生局基金资助 (编号 :0 448)
