[Multifunctional nano-vector for gene delivery into human adipose derived mesenchymal stem cells and in vitro cellular magnetic resonance imaging].
OBJECTIVE
To examine the feasibility and efficacy of using superparamagnetic iron oxide nanoparticles coated with polyethylene glycol-grafted polyethylenimine (PEG-g-PEI-SPION) as a carrier for gene delivery into human adipose derived mesenchymal stem cells (hADMSCs) and in vitro cellular magnetic resonance imaging (MRI).


METHODS
PEG-g-PEI-SPION was synthesized as previously reported. Gel electrophoresis was performed to assess the pDNA condensation capacity of PEG-g-PEI-SPION. The particle size and zeta potential of PEG-g-PEI-SPION/pDNA complexes were determined by dynamic light scattering. Cytotoxicity of PEG-g-PEI-SPION was evaluated by CCK-8 assay with hADMSCs. Gene transfection efficiency of PEG-g-PEI-SPION in hADMSCs was quantified by flow cytometry. The cellular internalization of PEG-g-PEI-SPION/pDNA nanocomplexes was studied by confocal laser scanning microscopy and Prussian blue staining. MRI function of PEG-g-PEI-SPION was studied by in vitro cellular MRI scanning.


RESULTS
PEG-g-PEI-SPION condensed pDNA to form stable complexes of 80-100 nm in diameter and showed low cytotoxicity in hADMSCs. At the optimal N/P ratio of 20, PEG-g-PEI-SPION/pDNA obtained the highest transfection efficiency of 22.8% ± 3.6% in hADMSCs. And it was higher than that obtained with lipofectamine 11.2% ± 2.6% (P < 0.05). Furthermore, hADMSCs labeled with PEG-g-PEI-SPION showed sensitive low signal intensity on MRI T2-weighted images in vitro.


CONCLUSIONS
PEG-g-PEI-SPION is an efficient and MRI-visible nano-vector for gene delivery into hADMSCs.
Laboratory of Molecular Imaging, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China.
Pengfei
P
Pang
Bing
B
Li
Xiaojun
X
Hu
Zhuang
Z
Kang
Shouhai
S
Guan
Faming
F
Gong
Xiaochun
X
Meng
Dan
D
Li
Mingsheng
M
Huang
Email: shanhong@mail.sysu.edu.cn.
Hong
H
Shan
D000273:Adipose Tissue / Q000166:cytology; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008279:Magnetic Resonance Imaging / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D053758:Nanoparticles; D014162:Transfection