GFP gene transfection of dendritic cells mediated by recombinated adeno-associated virus.
- Author:
Xiao-ying ZHOU
1
;
Ming JI
;
Jun-yan LIU
;
Meng-qun TAN
Author Information
1. Experimental Hematology Laboratory, Department of Physiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Antigens, CD34;
Bone Marrow Cells;
cytology;
Cell Differentiation;
Dendritic Cells;
cytology;
Dependovirus;
genetics;
Genetic Vectors;
Green Fluorescent Proteins;
genetics;
Hematopoietic Stem Cells;
cytology;
Humans;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2007;32(5):742-746
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the transfection condition of Type 2 recombinant adeno-associated virus ( rAAV2) in human dendritic cells(DCs) which were induced from the bone marrow CD34+ hematopoietic stem /progenitor cells.
METHODS:CD34+ hematopoietic stem /progenitor cells were purified from the bone marrow mononuclear cells by immunomagnetic beads, and the cells were cultured with IL-4 and GM-CSF and maturated by TNF-alpha on the 5th day. The rAAV2 /GFP was transfected into the induced cells at different time.The DCs were identified by electronic microscope. The expression of GFP was evaluated by flow cytometry and fluorescence microscope.
RESULTS:The DCs were induced successfully. The typical morphologic characteristics of DCs were observed under the light microscope and transmission electronic microscope, and the typical phenotypes of DCs could be detected by flow cytometry. The expression rate of GFP gene on the 3rd day, the 5th day and after adding TNF-alpha was 0.45%, 13.54%, and 0.25%, respectively.
CONCLUSION:DCs can be induced from the human bone marrow CD34+ hematopoietic stem /progenitor cells, and infected with the rAAV2 /GFP successfully. The longer the induction time of DCs, the higher the transfection efficiency of DC. The transfection efficiency of immature DC is higher than that of mature DC.
