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J Gene Med. 2014 Mar-Apr;16(3-4):84-96. doi: 10.1002/jgm.2762.

Optimizing cationic and neutral lipids for efficient gene delivery at high serum content.

Author information

1
Department of Materials, Department of Physics, and Molecular, Cellular & Developmental Biology Department, University of California at Santa Barbara, California 93106, USA.
2
Institute of Physics, Academia Sinica, Taipei 11529, Taiwan.
3
National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan.
4
Department of Electrophysics, National Chiao-Tung University, Hsinchu 30010, Taiwan.
#
Contributed equally

Abstract

BACKGROUND:

Cationic liposome (CL)-DNA complexes are promising gene delivery vectors with potential application in gene therapy. A key challenge in creating CL-DNA complexes for application is that their transfection efficiency (TE) is adversely affected by serum. In particular, little is known about the effects of a high serum content on TE, even though this may provide design guidelines for application in vivo.

METHODS:

We prepared CL-DNA complexes in which we varied the neutral lipid [1,2-dioleoyl-sn-glycerophosphatidylcholine, glycerol-monooleate (GMO), cholesterol], the headgroup charge and chemical structure of the cationic lipid, and the ratio of neutral to cationic lipid; we then measured the TE of these complexes as a function of serum content and assessed their cytotoxicity. We tested selected formulations in two human cancer cell lines (M21/melanoma and PC-3/prostate cancer).

RESULTS:

In the absence of serum, all CL-DNA complexes of custom-synthesized multivalent lipids show high TE. Certain combinations of multivalent lipids and neutral lipids, such as MVL5(5+)/GMO-DNA complexes or complexes based on the dendritic-headgroup lipid TMVLG3(8+) exhibited high TE both in the absence and presence of serum. Although their TE still dropped to a small extent in the presence of serum, it reached or surpassed that of benchmark commercial transfection reagents, particularly at a high serum content.

CONCLUSIONS:

Two-component vectors (one multivalent cationic lipid and one neutral lipid) can rival or surpass benchmark reagents at low and high serum contents (up to 50%, v/v). We propose guidelines for optimizing the serum resistance of CL-DNA complexes based on a given cationic lipid.

KEYWORDS:

cationic liposomes; gene delivery; glycerol monooleate; multivalent cationic lipid; serum

PMID:
24753287
PMCID:
PMC4051313
DOI:
10.1002/jgm.2762
[Indexed for MEDLINE]
Free PMC Article

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