Human skin fibroblasts transdution with a pseudovirus in assocation to Magnetofection transfection reagent

Human dermal fibfoblasts transduction was achieved using pseudovirus containing lentiviral expression with the Magnetofection method from OZ Biosciences.

This paper shows the efficiency of the Magnetofection technology from OZ Biosciences to enhance infection and transduce human skin fibroblasts with pseudovirus.

article reference: J Biol Chem. 2013 Jun 27.

Niemann-Pick Type C2 deficiency in human fibroblasts confers robust and selective activation of prostaglandin E2 biosynthesis.
Frolov A, Dong H, Jiang M, Yang L, Cook EC, Matnani R, Hammock BD, Crofford LJ.

Abstract

Activated fibroblasts, also known as myofibroblasts, are mediators of several major human pathologies including proliferative fibrotic disorders, invasive tumor growth, rheumatoid arthritis and atherosclerosis. We have previously identified Niemann-Pick Type C2 (NPC2) protein as a negative regulator of fibroblast activation. (Csepeggi, C. et al. 2011. J. Biol. Chem. 286:2078). Here we report that NPC2-deficiency leads to a dramatic up-regulation of the arachidonic acid (AA) metabolic pathway in human fibroblasts. The major enzymes in this pathway, cPLA2 type IVA, COX-2, and mPGES-1 were dramatically upregulated at both the transcriptional and translational levels. The specific phenotypic changes resulted in a more than 10-fold increase in the production and secretion of a key modulator of inflammation and immunity, prostaglandin E2 (PGE2). More importantly, AA metabolome profiling by liquid chromatography / tandem mass-spectrometry (LC/MS/MS) revealed the very specific nature of PGE2 up-regulation as the other analyzed AA metabolites derived from the COX-2, CYP450, 5/15-LOX, and non-enzymatic oxidative pathways were mostly down-regulated. Blocking activity of cPLA2 efficiently suppressed expression of inflammatory cytokines, IL-1β and IL-6, thereby identifying cPLA2 as an important regulator of the inflammatory program in NPC2-null cells. Altogether, these studies highlight NPC2 as a specific regulator of AA metabolism and inflammation that suggests potential for NPC2 protein or its related signaling in the treatment of inflammatory diseases characterized by the presence of activated fibroblasts.

OZ Biosciences provides several magnetic nanoparticles formulations optimized for increasing any kind of virus infection and transduction both in vitro and in vivo.