CAJ | 학술논문

目的通过实验验证一个体外大量扩增晚期内皮祖细胞( EPC)的方法.方法先预铺经辐射后的滋养层细胞(晚期EPC或脐带脐静脉内皮细胞),而后植入分离的人外周血单个核细胞,计数并对比培养21 d后获得的晚期EPC克隆数目,并检测目的细胞免疫表型,应用基质胶( matrigel)鉴定其体外管状结构形成能力.结果 21 d时在预铺的两种滋养层细胞培养条件下获得的晚期EPC克隆数目均比无滋养层细胞的培养方法高20倍(40.0±3.9、39.3 ±3.1比2.0±1.3,均P＜0.05)；且细胞克隆出现的早.目的细胞表达CD31、CD34、内皮型一氧化氮合酶(eNOS)、血管内皮细胞生长因子受体Flt-1、CD146的2个表型(P1H12、Sendo)、血管内皮钙黏蛋白(VEcadherin)、CD117,且具有能在体外与matrigel形成管状结构的功能.结论通过提供滋养层细胞的方法可以大量扩增晚期EPC,提示微环境在晚期EPC扩增中起重要作用.
목적 통과실험험증일개체외대량확증만기내피조세포( EPC)적방법.방법 선예포경복사후적자양층세포(만기EPC혹제대제정맥내피세포),이후식입분리적인외주혈단개핵세포,계수병대비배양21 d후획득적만기EPC극륭수목,병검측목적세포면역표형,응용기질효( matrigel)감정기체외관상결구형성능력.결과 21 d시재예포적량충자양층세포배양조건하획득적만기EPC극륭수목균비무자양층세포적배양방법고20배(40.0±3.9、39.3 ±3.1비2.0±1.3,균P＜0.05)；차세포극륭출현적조.목적세포표체CD31、CD34、내피형일양화담합매(eNOS)、혈관내피세포생장인자수체Flt-1、CD146적2개표형(P1H12、Sendo)、혈관내피개점단백(VEcadherin)、CD117,차구유능재체외여matrigel형성관상결구적공능.결론 통과제공자양층세포적방법가이대량확증만기EPC,제시미배경재만기EPC확증중기중요작용.
Objective To validate a novel method of expanding late endothelial progenitor cells (EPC) in vitro.Methods We cultured mononuclear cells (MNC) from human peripheral blood on the plate with the feeder layer cells,i.e.irradiated late EPC or human umbilical vein endothelial cells.After 21days,the numbers of late EPC colonies were counted separately. And the surface antigen of late EPC was detected by fluorescence-activated cell sorter (FACS) and their in vitro ability of forming vascular structure examined by matrigel.Results Both colony numbers of late EPC with feeder layer cell culturing were over 20 times than those without (40.0 ±3.9,39.3 ±3.1 vs 2.0 ± 1.3,both P ＜0.05).And the former's late EPC colony appeared earlier.The late EPC expressed CD31,CD34,eNOS,Flt-1,P1H12,Sendo,VE cadherin and CD117. And vascular structures were discerned.Conclusions The method of feeder layer cells may vastly expand the quantity of late EPC. And microenvironment plays an important role in its expansion.