Palmitic acid substitution on cationic polymers for effective delivery of plasmid DNA to bone marrow stromal cells.
Nonviral gene carriers are actively explored in gene therapy due to safety concerns of the viral carriers. To design effective gene carriers for modification of bone marrow stromal cells (BMSC), an important cell phenotype for clinical application of gene therapy, cationic polymers polyethyleneimine (PEI), and poly-L-Lysine (PLL) were substituted with palmitic acid (PA) via amide linkages. Depending on the reaction conditions, PEI and PLL was substituted with 2.2-5.2 and 13.4-16.2 PA per polymer chain. The PA substituted polymers displayed slightly lower binding efficiency towards a plasmid containing Enhanced Green Fluorescent Protein (pEGFP) in an agarose gel binding assay. The cell binding of PLL-PA, but not PEI-PA, was particularly enhanced, resulting in higher percentage of the cells displaying a significant polymer uptake. pEGFP delivery into the BMSC was also significantly increased with the PLL-PA (vs. PLL), but not PEI-PA (vs. PEI). The transfection efficiency of PLL-PA was significantly higher ( approximately fivefold) than the unmodified polymer. We conclude that PA substitution on PLL provides an effective carrier for transfection of primary cells derived from the bone marrow.
Incani
Vanessa
V
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Tunis
Emily
E
Clements
Basak Acan
BA
Olson
Cori
C
Kucharski
Cezary
C
Lavasanifar
Afsaneh
A
Uludag
Hasan
H
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D001854:Bone Marrow Cells / Q000378:metabolism*; D002412:Cations / Q000737:chemistry; D016503:Drug Delivery Systems; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D066298:In Vitro Techniques; D008422:Materials Testing; D019308:Palmitic Acid / Q000737:chemistry; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics; D017154:Stromal Cells / Q000378:metabolism; D014162:Transfection
D001672:Biocompatible Materials; D002412:Cations; D011108:Polymers; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011107:Polylysine; D019308:Palmitic Acid; D011094:Polyethyleneimine