[Transduction efficiency of recombinant adeno-associated virus 2 in human bone marrow CD34+ hematopoietic stem/progenitor cells and mesenchyme stem cells].
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.
Sun
Xiao-juan
XJ
Blood Physiology Laboratory, Department of Physiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Wang
Feng
F
Tan
Meng-qun
MQ
Lu
Guang-xiu
GX
D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology; D000229:Dependovirus / Q000235:genetics*; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000378:metabolism* / Q000821:virology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism* / Q000821:virology; D011995:Recombination, Genetic; D014161:Transduction, Genetic
D018952:Antigens, CD34; D049452:Green Fluorescent Proteins