Liposome-mediated gene transfer to fetal human ventral mesencephalic explant cultures (original) (raw)
Related papers
Non-virally mediated gene transfer into human central nervous system precursor cells
Molecular Brain Research, 1996
Lipofectamine-based transfection was used as a method of choice to deliver the bacterial β-galactosidase gene into human central nervous system (CNS) precursor cells. We achieved a transfection efficiency of 7.4%. β-Galactosidase expressing cells were shown to display both neuronal and glial phenotypes. We also delivered the temperature sensitive allele of SV40 Large-T antigen and obtained a high level of expression of the immortalizing oncoprotein in the cells. Colonies of Large-T antigen immunoreactive cells were indeed visible 10 days after transfection.
Liposomal and Viral Vectors for Gene Therapy of the Central Nervous System
Current Drug Targets - Cns & Neurological Disorders, 2005
Due to the presence of the blood-brain barrier, the central nervous system (CNS) is not easily accessible to systemically delivered macromolecules with therapeutic activity such as growth factors, cytokines or enzymes. Therefore, the expression of exogenously administered genes in the brain has been proposed for a wide variety of inherited and acquired diseases of the CNS, for which classical pharmacotherapy is unavailable or not easily applicable. Gene therapy to the CNS has been the target of a great number of studies aiming at finding a viable therapeutic strategy for the treatment of neurological disorders. This approach has already been used as a promising tool for brain protection and repair from neuronal insults and degeneration in several animal models, and is currently being applied in clinical trials.
Methods for Gene Transfer to the Central Nervous System
Advances in Genetics, 2014
Gene transfer is an increasingly utilized approach for research and clinical applications involving the central nervous system (CNS). Vectors for gene transfer can be as simple as an unmodified plasmid, but more commonly involve complex modifications to viruses to make them suitable gene delivery vehicles. This chapter will explain how tools for CNS gene transfer have been derived from naturally occurring viruses. The current capabilities of plasmid, retroviral, adenoassociated virus, adenovirus, and herpes simplex virus vectors for CNS gene delivery will be described. These include both focal and global CNS gene transfer strategies, with short-or longterm gene expression. As is described in this chapter, an important aspect of any vector is the cisacting regulatory elements incorporated into the vector genome that control when, where, and how the transgene is expressed.
Folia morphologica, 2005
CNS gene transfer could provide new approaches to the modelling of neurodegenerative diseases and devising potential therapies. One such disorder is Parkinson's disease (PD), in which dysfunction of several different metabolic processes has been implicated. Here we review the literature on gene transfer systems based on herpes simplex virus type 1 (HSV-1) and non-viral polyethyleneimine (PEI) and calcium phosphate nanoparticle methods. We also assess the usefulness of various CNS gene delivery methods and present some of our own data to exemplify such usefulness. Our data result from vectors stereotaxically introduced to the substantia nigra (SN) of adult rats and evaluated 1 week and/or 1 month post injection using histochemical methods to assess recombinant ss-galactosidase enzyme activity. Gene transfer using PEI or calcium phosphate-mediated transfections was observed for both methods and PEI was comparable to that of HSV-1 amplicon. Our data show that the amplicon delivery ...
Lipospermine-mediated gene transfer technique into murine cultured cortical cells
Journal of Neuroscience Methods, 1997
In order to transfer exogenous DNA into embryonic cortical cells, we have chosen a transfection technique using a synthetic lipospermine (dipalmitoylphosphatidylethanolamylspermine, DPPES) which complexes DNA molecules and allows their penetration into the intracellular compartment. The procedure was optimized after testing several parameters: DPPES/DNA ratio, incubation time, kinetics of transgene expression, and growth medium. The protocol was achieved by following the expression of the E. coli LacZ reporter gene under the control of the cytomegalovirus promoter. The lipopolyamine-mediated transfection is efficient for terminally differentiated cells, since we routinely obtained transfection efficiencies of 30% for neurons. © 1997 Elsevier Science B.V.
Global non-viral gene transfer to the primate brain following intravenous administration
Molecular Therapy, 2003
Expression plasmids encoding either luciferase or -galactosidase were encapsulated in the interior of an "artificial virus" comprised of an 85 nm pegylated immunoliposome, which was targeted to the rhesus monkey brain in vivo with a monoclonal antibody (MAb) to the human insulin receptor (HIR). The HIRMAb enables the liposome carrying the exogenous gene to undergo transcytosis across the blood-brain barrier and endocytosis across the neuronal plasma membrane following intravenous injection. The level of luciferase gene expression in the brain was 50-fold higher in the rhesus monkey as compared to the rat. Widespread neuronal expression of the -galactosidase gene in primate brain was demonstrated by both histochemistry and confocal microscopy. This approach makes feasible reversible adult transgenics in 24 hours.