title: Transfecting the hard-to-transfect lymphoma/leukemia cells using a simple cationic polymer nanocomplex.
abstract: 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 200nm 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 (>7kDa) 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.
authors.affiliation: Department of Pathology and Genomic Medicine, The Methodist Hospital and The Methodist Hospital Research Institute, Houston, TX 77030, USA.
authors.forename: Nianxi
authors.initials: N
authors.lastname: Zhao
authors.forename: Jianjun
authors.initials: J
authors.lastname: Qi
authors.forename: Zihua
authors.initials: Z
authors.lastname: Zeng
authors.forename: Parag
authors.initials: P
authors.lastname: Parekh
authors.forename: Chung-Che
authors.initials: CC
authors.lastname: Chang
authors.forename: Ching-Hsuan
authors.initials: CH
authors.lastname: Tung
authors.forename: Youli
authors.initials: Y
authors.lastname: Zu
mesh_terms: D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007938:Leukemia; D008223:Lymphoma; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D011108:Polymers / Q000008:administration & dosage* / Q000737:chemistry; D014162:Transfection / Q000379:methods*
chemical_list: D011108:Polymers; C507253:poly(beta-amino ester); D049452:Green Fluorescent Proteins; D004247:DNA