Journal: Neuropathology and Applied Neurobiology

Article Title: Development of a culture system to induce microglia-like cells from haematopoietic cells

doi: 10.1111/nan.12086

Figure Lengend Snippet: Differentiation assays of LN − cells. ( A ) Immunocytology of mouse LN − cells cultured on murine astrocytes with or without M-CSF or IL-34. Bar: 50 μm. ( B ) Dose–response relationship between M-CSF/IL-34 and the number of ML cells differentiated from LN − cells (cell number per mm 2 ). * P < 0.05 as compared with untreated cells. ( C ) Quantitative analysis of small round-shaped cells and large flat cells differentiated from LN − cells (cell number per mm 2 ). ( D ) Morphological characteristics of LN − cells differentiated with M-CSF or IL-34. Bar: 10 μm. ( E , F ) Quantitative analysis ( E ) and immunocytology ( F ) of mouse monocytes cultured on murine astrocytes in the absence or presence of M-CSF or IL-34, Bar: 50 μm. ( G , H ) Fluorescent microscopic images ( G ) and quantitative analysis ( H ) of LN − cells cultured on astrocytes with or without anti-IL-34 antibody. IL-34 neutralization resulted in the significant reduction of round cells. * P < 0.05 as compared with cells treated by control IgG. Bar: 50 μm. ( I ) Intracellular staining of IL-34 of mouse primary mixed glial cells. Some GFAP-positive astrocytes showed granular staining in cytoplasm (arrowhead), whereas other astrocytes were not stained with anti-IL-34 Ab (arrow), Bar: 20 μm. Data are presented as mean ± SD. * P < 0.05, ** P < 0.005, *** P < 0.00005.

Article Snippet: Antibodies (Abs) against mouse CD3, CD4, CD5, CD8α, CD11b, B220, Gr-1, TER-119, TREM2 and IL-34 (for neutralization) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Neutralization, Staining