TY - JOUR
T1 - Transfecting the hard-to-transfect lymphoma/leukemia cells using a simple cationic polymer nanocomplex
AU - Zhao, Nianxi
AU - Qi, Jianjun
AU - Zeng, Zihua
AU - Parekh, Parag
AU - Chang, Chung Che
AU - Tung, Ching Hsuan
AU - Zu, Youli
N1 - Funding Information:
This study was supported by National Cancer Institute grant RO1CA151955 and the Collaborative Research Fund from the Virginia and L.E. Simmons Family Foundation (to Dr. Youli Zu).
PY - 2012/4/10
Y1 - 2012/4/10
N2 - Although the development of gene delivery systems via non-viral-mediated methods is advancing rapidly, it remains a challenge to deliver plasmids into hard-to-transfect cells, such as lymphoma/leukemia cells. To develop an efficient transfection method, we formulated a simple nanocomplex by incorporating poly β-amino ester (PBAE) polymers with plasmid DNAs containing a GFP reporter gene. The formed PBAE-plasmid nanocomplexes are approximately 200 nm in diameter and stable under physiological conditions, but become rapidly biodegradable when pH decreases < 7.0. Cultured lymphoma/leukemia cells were used for transfection assays and resultant gene delivery rates were determined by quantifying GFP expression. Exposure of cells to the nanocomplexes composed of fractioned PBAE (> 7 kDa) resulted in GFP expression in 3% of cells, similar to that mediated by the standard Lipofectamine method. However, with polybrene pre-treatment, the nanocomplex could achieve GFP expression in up to 32% of lymphoma/leukemia cells, an 8-fold increase over that mediated by Lipofectamine. These findings demonstrated a simple, efficient method for in vitro gene delivery into hard-to-transfect cells. The nanocomplexes are biodegradable and have minimal cytotoxicity, suggesting the potential use for in vivo gene delivery.
AB - Although the development of gene delivery systems via non-viral-mediated methods is advancing rapidly, it remains a challenge to deliver plasmids into hard-to-transfect cells, such as lymphoma/leukemia cells. To develop an efficient transfection method, we formulated a simple nanocomplex by incorporating poly β-amino ester (PBAE) polymers with plasmid DNAs containing a GFP reporter gene. The formed PBAE-plasmid nanocomplexes are approximately 200 nm in diameter and stable under physiological conditions, but become rapidly biodegradable when pH decreases < 7.0. Cultured lymphoma/leukemia cells were used for transfection assays and resultant gene delivery rates were determined by quantifying GFP expression. Exposure of cells to the nanocomplexes composed of fractioned PBAE (> 7 kDa) resulted in GFP expression in 3% of cells, similar to that mediated by the standard Lipofectamine method. However, with polybrene pre-treatment, the nanocomplex could achieve GFP expression in up to 32% of lymphoma/leukemia cells, an 8-fold increase over that mediated by Lipofectamine. These findings demonstrated a simple, efficient method for in vitro gene delivery into hard-to-transfect cells. The nanocomplexes are biodegradable and have minimal cytotoxicity, suggesting the potential use for in vivo gene delivery.
KW - β amino ester polymers
KW - Cell transfection
KW - Gene delivery
KW - Hard-to-transfect
KW - Leukemia/lymphoma
KW - Polybrene
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U2 - 10.1016/j.jconrel.2012.01.007
DO - 10.1016/j.jconrel.2012.01.007
M3 - Article
C2 - 22269663
AN - SCOPUS:84862795899
VL - 159
SP - 104
EP - 110
JO - Journal of Controlled Release
JF - Journal of Controlled Release
SN - 0168-3659
IS - 1
ER -