The primary glial cultures prepared with this method have the great advantage of being devoid of neurons, enabling the analysis of astrocytes and oligodendrocytes separately. This culture model has been extensively used to advance our knowledge in the field of glial biology in normal conditions and ...
Cell culture Primary cultured neurons and astrocytes were prepared from the mesencephalon and striata, respectively, of SD rat embryos at 15 days of gestation using a method described previously33. Dissected mesencephalons and striata were cut into small pieces with scissors and incubated in 0.125% ...
An MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed according to the manufacturer protocol. Primary neuronal cells were seeded in PDL-laminin coated 96-well culture plates at a density of 1 × 104cells per well in triplicates and grown until 70–8...
Graph shows quantification of GFAP+ cell area in culture. Data are whisker boxes (min to max, center line indicating median). **p < 0.01 (two-tailed Mann–Whitney’s test).eLeft, Fluorescence micrographs of immunostained primary astrocytes (GFAP, desaturated...
Primary astrocytes express low levels of these receptors and are unresponsive to Thy-1; thus, inflammation and astrocyte reactivity might be necessary for Thy-1-induced responses. Methods Wild-type rat astrocytes (TNF-activated) or from human SOD1G93A transgenic mice (a neurodegenerative disease ...
Cell culture Primary cultures of cortical astrocytes were prepared from the cortices of neonatal 1-day-old C57BL/6 mice using methods described in a previous stuciefly, cortices were dissected from the brains, and the meninges were carefully removed [19]. Then, the cortices were dissociated into...
Mn activated the transcription factor YY1, leading to decreased GLT-1/GLAST mRNA/protein levels in astrocyte culture (21), indicating a potential role for YY1 in Mn-induced neurotoxicity. Accordingly, the results from the present study confirm that deletion of astrocytic YY1 attenuated Mn-induced ...
protocol as compared to primary brain tissue samples. After binding to antibody-coated plates, the cells of interest were either collected via trypsinization for subsequent culture in 2D or directly scraped off the plate to extract RNA for transcriptional profiling....
bEnd.3 cells were labeled with CFSE, and primary astrocytes were grown on the bottom of the culture dish. E, F are western blot analysis of FPN1 levels in groups treated as in C and D. The relative expression levels were normalized to β-actin levels and expressed as the mean ± ...
Bright PrPSc puncta (red) were detectable within β 3-tubulin-positive neuronal cell bodies (green) suggesting transfer of infectivity could occur in primary culture as well. Only the upper z-stacks of the image are shown for clarity to reduce signal from astrocytes and focus on neuronal ...