CAJ | 학술논문

目的从脐带中分离内皮祖细胞(EPCs),考察其体外增殖、基因转染绿色荧光蛋白质粒的行为.方法以酶消化法从脐带中分离出内皮祖细胞,并通过流式细胞仪和共聚焦显微镜对内皮祖细胞进行鉴定,以Lipofectamine 2000为转染试剂考察了内皮祖细胞转染绿色荧光蛋白质粒的行为.结果从脐带中分离培养的内皮祖细胞在第9天形成了典型的内皮细胞集落,流式细胞仪分析结果显示CD133和激酶插入区受体(KDR)的含量均有所提高,并具有内皮祖细胞结合异硫氰酸荧光素(FITC)标记的荆豆凝聚素1(FITC-UEA-1)和吞噬DiI标记的低密度脂蛋白(DiI-ac-LDL)的功能,能较好地表达绿色荧光蛋白.结论从脐带中分离的内皮祖细胞体外在适当的培养条件下,可增殖、诱导分化为内皮细胞,并能较好地表达外源基因,是基因与细胞治疗理想的载体.
목적 종제대중분리내피조세포(EPCs),고찰기체외증식、기인전염록색형광단백질립적행위.방법 이매소화법종제대중분리출내피조세포,병통과류식세포의화공취초현미경대내피조세포진행감정,이Lipofectamine 2000위전염시제고찰료내피조세포전염록색형광단백질립적행위.결과 종제대중분리배양적내피조세포재제9천형성료전형적내피세포집락,류식세포의분석결과현시CD133화격매삽입구수체(KDR)적함량균유소제고,병구유내피조세포결합이류청산형광소(FITC)표기적형두응취소1(FITC-UEA-1)화탄서DiI표기적저밀도지단백(DiI-ac-LDL)적공능,능교호지표체록색형광단백.결론 종제대중분리적내피조세포체외재괄당적배양조건하,가증식、유도분화위내피세포,병능교호지표체외원기인,시기인여세포치료이상적재체.
Objective To isolate and identify endothelial progenitor cells (EPCs) from human umbilical cord,and to study the cell proliferation and gene transfection of green fluorescent protein plasmid in vitro.Methods EPCs were isolated from human umbilical cord in enzyme digestion method.The biological characteristics of EPCs were identified by flow cytometry and laser confocal microscope.The enhanced green fluorescent protein (EGFP) gene transfection mediated by EPCs was investigated using Lipofectamine 2000 as transfection reagent.Results Endothelial progenitor cells isolated from umbilical cord formed typical endothelial cell colony 9 days later.These cells displayed an improved positive expression of CD133 and kinase insert domain receptor (KDR).The endotheliallineage characteristics of expanded cells were confirmed by fluorescein isothiocyanate (FITC)-UEA-1 binding and DiI-ac-LDL uptake assay with the aid of laser confocal microscope.The transfection results demonstrated high expression of EGFP taking EPCs as host cell.Conclusion Endothelial progenitor cells isolated from umbilical cord can be propagated and induced to differentiate into endothelial cells in the appropriate culture conditions.EPCs demonstrated to be an ideal carrier for gene and cell therapy.