Chitosan lactate as a nonviral gene delivery vector in COS-1 cells.
The purpose of this research was to evaluate chitosan lactate (CL) of different molecular weights (MWs) as a DNA complexing agent for its efficiency in transfecting COS-1 cells (green monkey fibroblasts) and its effect on cell viability compared with polyethylenimine (PEI), a commercially available cationic polymer. CL and chitosan base dissolved in dilute acetic acid (chitosan acetate [CA]) of different MWs (20, 45, 200, 460 kDa) and N/P ratios (2:1, 4:1, 8:1, 12:1, 24:1) formed complexes with pSV beta-galactosidase plasmid DNA. The complexes were characterized by agarose gel electrophoresis and investigated for their ability to transfect COS-1 cells compared with PEI. Additionally, the effect of CL on the viability of COS-1 cells was investigated using 3-(4,5-dimethyliazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The binding of CL/DNA and CA/DNA was dependent on chitosan MWs. The N/P ratio of CL to completely form the complex with the DNA was higher than that of CA. Both CL and CA were comparable in transfection efficiencies at an N/P ratio of 12:1, but less efficient than PEI (P < .05). The cell viability in the presence of CL and CA at all MWs was over 90%, whereas that of PEI-treated cells was approximately 50%. These results suggest the advantage of CL for in vitro gene transfection, with the ease of preparation of polymer/DNA complexes and low cytotoxicity.
Weecharangsan
Wanlop
W
Faculty of Pharmacy, Silpakorn University, Nakhonpathom 73000, Thailand.
Opanasopit
Praneet
P
Ngawhirunpat
Tanasait
T
Rojanarata
Theerasak
T
Apirakaramwong
Auayporn
A
D000818:Animals; D019556:COS Cells; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000008:administration & dosage* / Q000737:chemistry; D002522:Chlorocebus aethiops; D004247:DNA / Q000008:administration & dosage*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D019344:Lactic Acid / Q000737:chemistry; D011094:Polyethyleneimine / Q000008:administration & dosage*; D014162:Transfection
D019344:Lactic Acid; D011094:Polyethyleneimine; D004247:DNA; D048271:Chitosan