Specific Transfection of Inflamed Brain by Macrophages: A New Therapeutic Strategy for Neurodegenerative Diseases
Figure 7
Transfection of Cath.A neurons by GFP-transfected macrophages.
RAW 264.7 macrophages were transfected with GFP pDNA, cultured in complete media for three days, and then added to Cath.A neurons. To distinguish between the cell types, macrophages were stained with CD11b Ab (Alexa 647). GFP levels in neurons were assessed by FACS as mean fluorescence ± SEM (N = 4). A: The representative FACS plots demonstrating GFP transfer into Cath.A neurons; B: Quantification of GFP levels in macrophages alone (black diamonds), and in co-culture of neurons and macrophages (white squares). GFP expression levels in neurons co-cultured with transfected macrophages increased over 5–12 days. At the same time, protein expression in macrophages at days 5–12 was already diminished, suggesting that along with GFP, its genetic material (pDNA and RNA) was transferred from transfected macrophages into neurons, where the encoded protein (GFP) was synthesized de novo. Statistical significance shown by asterisk (p<0.05) was calculated by a one-way ANOVA. The bar: 10 µm.