Experimental study of human umbilical cord blood derived stromal cells transfected with recombinant adenoviral vector co-expressing VCAM-1 and GFP.
- Author:
Xi ZHANG
1
;
Ying-Jian SI
;
Xing-Hua CHEN
;
Yao LIU
;
Li GAO
;
Lei GAO
;
Xian-Gui PENG
;
Qing-Yu WANG
Author Information
1. Department of Hematology, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Fetal Blood;
cytology;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
genetics;
metabolism;
Humans;
RNA, Messenger;
genetics;
metabolism;
Recombinant Proteins;
genetics;
metabolism;
Stromal Cells;
cytology;
Transfection;
Vascular Cell Adhesion Molecule-1;
genetics;
metabolism
- From:
Journal of Experimental Hematology
2008;16(3):598-604
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the effect of vcam-1 gene-modified human umbilical cord blood derived stromal cells (CBDSCs) on hematopoietic regulation so as to establish the experimental foundation for further study. The target gene vcam-1 was cloned into the shuttle plasmid with the report gene GFP. The recombinant shuttle plasmid was transformed into BJ5183 bacteria to recombine with backbone vector pAdeasy-l, and the recombinant adenoviral vector ad-vcam-1-gfp was confirmed after transfection with CBDSCs. The results indicated that two fragments of about 9 kb and 2 kb were obtained after digestion of recombinant plasmid pAdTrack-vcam-1 with NotIand XhoI, and single fragment of 600 bp was obtained after amplification with PCR; two fragments of about 31 kb and 4 kb were obtained after digestion of recombinant plasmid pad-vcam-1-gfp with PacI, which suggested a successful homologous recombination. The expression of vcam-1 gene in ad-vcam-1-gfp transfected CBDSCs could be detected by immunocytochemistry, RT-PCR and fluorescent microscopy. It is concluded that the recombinant adenoviral vector ad-vcam-1-gfp has been constructed successfully, and the expression of vcam-1 is up-regulated in CBDSCs transfected by gene ad-vcam-1-gfp.
