Fig. 4
From: The pathogenic role of c-Kit+ mast cells in the spinal motor neuron-vascular niche in ALS
c-Kit+ mast cell precursors engraft into the degenerating spinal cord of SOD1G93A mice following i.v. administration. A The scheme shows the isolation and culture of mast cells precursors from the bone marrow of Non-Tg mice and the protocol followed for the i.v. injection c-Kit+ mast cell precursors stained with the CFSE cell tracer in Non-Tg and symptomatic SOD1G93A littermates at 150 days. B Left panels show representative 3D confocal images of c-Kit immunostaining of mast cells precursors. Right panels show the flow cytometry density plot analysis c-Kit+ (red—left panel) precursors stained with CFSE+ (green—right panel). C Representative confocal images showing infiltration of c-Kit+/CFSE+ mast cells precursors (magenta arrows) into the symptomatic SOD1G93A spinal cord parenchyma following 48 h after i.v. administration. c-Kit+/CFSE+ cells engrafted the parenchyma in the surroundings of blood vessels assessed by EB systemically perfused after euthanasia (white). Magenta squares show high magnifications 3D reconstructions of typical c-Kit+/CFSE+ precursor cells. D Representative confocal image of the co-expression of c-Kit and CFSE in the spinal cord of i.v. injected Non-Tg mice. None of few cells were observed in the spinal cord parenchyma of controls. E The graph shows the quantitative analysis of c-Kit+/CFSE+cells in Non-Tg and SOD1G93A mice. Quantitative data are expressed as mean ± s.e.m. Data were analyzed by two-tailed Mann–Whitney test, with***p < 0.001 considered significant. Scale bars: 10 μm (Aand B) and 5 μm in (A)