Research Article
The Comparative Utility of Viromer RED and Lipofectamine for
Transient Gene Introduction into Glial Cells
Sudheendra Rao,
1
Alejo A. Morales,
1
and Damien D. Pearse
1,2,3,4
1
he Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA
2
he Departments of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA
3
he Neuroscience Program, University of Miami Miller School of Medicine, Miami, FL 33136, USA
4
he Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL 33136, USA
Correspondence should be addressed to Sudheendra Rao; sudhee26@med.miami.edu
Received 24 April 2015; Revised 23 July 2015; Accepted 26 July 2015
Academic Editor: Eiry Kobatake
Copyright © 2015 Sudheendra Rao et al. his is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
he introduction of genes into glial cells for mechanistic studies of cell function and as a therapeutic for gene delivery is an expanding
ield. hough viral vector based systems do exhibit good delivery eiciency and long-term production of the transgene, the need
for transient gene expression, broad and rapid gene setup methodologies, and safety concerns regarding in vivo application still
incentivize research into the use of nonviral gene delivery methods. In the current study, aviral gene delivery vectors based upon
cationic lipid (Lipofectamine 3000) lipoplex or polyethylenimine (Viromer RED) polyplex technologies were examined in cell
lines and primary glial cells for their transfection eiciencies, gene expression levels, and toxicity. he transfection eiciencies of
polyplex and lipoplex agents were found to be comparable in a limited, yet similar, transfection setting, with or without serum across
a number of cell types. However, diferential efects on cell-speciic transgene expression and reduced viability with cargo loaded
polyplex were observed. Overall, our data suggests that polyplex technology could perform comparably to the market dominant
lipoplex technology in transfecting various cells lines including glial cells but also stress a need for further reinement of polyplex
reagents to minimize their efects on cell viability.
1. Introduction
Recent studies have challenged our notions on glia : neuron
interactions and the role that glia play in normal physiology
as well as in the pathology of disease [1–4]. hus we are
at the crossroads of reexamining our understanding of the
role of glia in the nervous system. Glial cells play important
functions in immune modulation and responses to injury
including scarring, axon guidance, and remyelination repair.
herefore, glial cells from both central (astrocytes, oligoden-
drocytes, and microglia) and peripheral (Schwann cells) ner-
vous systems are emerging as attractive gene therapy targets
in a range of neurological disorders and trauma [5, 6]. Genetic
manipulation of glia, to modify their expression of speciic
molecules, can thus signiicantly alter their molecular and
physiological reactions to the environment, providing a tool
for better understanding their function under pathological
conditions as well as novel therapeutic targets for neuropro-
tection and neurorepair [7–9]. hough viral delivery systems
remain at the forefront of gene therapeutic approaches, safety
concerns and costs remain signiicant issues. Furthermore,
the need for fast development times and transient expression
paradigms in vitro and in vivo for gene delivery applications
still incentivize research into the use of nonviral gene delivery
methods. Nonviral gene delivery methods have improved
enormously in recent years and can ofer integration-free
expression that is becoming more comparable to that of
viral vectors under certain experimental conditions [10]. In
targeting glial cells, nonviral genetic manipulation has been
performed by physical (ballistic labelling, magnetofection),
electrical (electroporation), or chemical methods (cationic
polymer, cationic lipid, or calcium phosphate) [11–15].
Despite signiicant research investigation with chem-
ical transfection formulations of cationic lipids (forming
Hindawi Publishing Corporation
BioMed Research International
Volume 2015, Article ID 458624, 10 pages
http://dx.doi.org/10.1155/2015/458624