Isolation, identification and EGFP gene transfection of endothelial progenitor cells derived from human umbilical cord
10.3760/cma.j.issn.1673-4181.2012.01.011
- VernacularTitle:人脐带内皮祖细胞的分离、鉴定及转染EGFP质粒的实验研究
- Author:
Dunwan ZHU
;
Liping SONG
;
Lanxia LIU
;
Xia DONG
;
Hailing ZHANG
;
Hai WANG
;
Xigang LENG
- Publication Type:Journal Article
- Keywords:
Endothelial progenitor cells;
Umbilical cord;
Gene transfection
- From:
International Journal of Biomedical Engineering
2012;35(1):42-45
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo 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.MethodsEPCs 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.ResultsEndothelial progenitor cells isolated from umbilical cord formed typical endothelial cell colony 9 days later.These cellsdisplayed 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.ConclusionEndothelial 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.
