Chitosan lactate as a nonviral gene delivery vector in COS-1 cells |
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Authors: | Wanlop Weecharangsan Praneet Opanasopit Tanasait Ngawhirunpat Theerasak Rojanarata Auayporn Apirakaramwong |
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Institution: | (1) Department of Biopharmacy, Faculty of Pharmacy, Silpakorn University, 73000 Nakhonpathom, Thailand |
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Abstract: | 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 β-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 ≈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.
Published: August 4, 2006 |
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Keywords: | chitosan chitosan lactate transfection efficiency gene delivery COS-1 cells |
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