Transduction efficiency of recombinant adeno-associated virus 2 in human bone marrow CD34+ hematopoietic stem/progenitor cells and mesenchyme stem cells.
- Author:
Xiao-juan SUN
1
;
Feng WANG
;
Meng-qun TAN
;
Guang-xiu LU
Author Information
1. Blood Physiology Laboratory, Department of Physiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Antigens, CD34;
metabolism;
Bone Marrow Cells;
cytology;
metabolism;
virology;
Dependovirus;
genetics;
Genetic Therapy;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Hematopoietic Stem Cells;
metabolism;
virology;
Humans;
Mesenchymal Stem Cells;
metabolism;
virology;
Recombination, Genetic;
Transduction, Genetic
- From:
Journal of Central South University(Medical Sciences)
2006;31(1):19-23
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the transduction efficiency of recombinant adeno-associated virus 2 ( rAAV2) in human bone marrow CD34+ hematopoietic stem/progenitor cells and mesenchyme stem cells.
METHODS:The rAAV2 containing green fluorescent protein genes (rAAV2/GFP) were constructed, packaged and purified. CD34+ hematopoietic stem/progenitor cells and mesenchyme stem cells were infected with the rAAV2/GFP. After transduction for 48 hours, the expression of GFP was detected under fluorescence microscope. Furthermore, the transduction efficiency of AAV transduced CD34+ with hydroxyurea (HU) pretreatment and that of untreated were compared.
RESULTS:GFP genes were expressed in 5.3% +/- 1.7% CD34+ cells. After pretreatment with HU, the expression of the GFP gene in CD34+ cells increased to 13.2% +/- 2.8%, and 23% +/- 3.6% mesenchyme stem cells expressed the GFP gene. Conclusion The transduction efficiency of mesenchyme stem cells is higher than that of CD34+ hematopoietic stem/progenitor cells. HU pretreatment can obviously increase the transduction efficiency of CD34+ hematopoietic stem/progenitor cells.
