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Figure 1. STEMprep™ Mouse Brain Dissociation Kit Achieves High Cell Viability and Yield

Mouse brain tissue was processed into single-cell suspension using the STEMprep™ Mouse Brain Dissociation Kit and the STEMprep Tissue Dissociator, an alternative automated system, or a manual dissociation method. (A) Viability of nucleated cells. (B) Yield of viable cells per whole brain tissue (325 - 395 mg). (C and D) Proportion and yield of CD45+ immune and CD45- non-immune cells, including CD11b+F4/80+ microglia, O4+ oligodendrocytes, ACSA-2+ astrocytes, and CD31+ endothelial cells. Viability, yield, and subset composition were assessed by flow cytometry. Brain samples were processed with protocol-specific density gradient media to remove myelin and cell debris. Red blood cells were lysed with ammonium chloride solution prior to subset analysis. Data are presented as mean ± SD (n = 21 - 36), * p < 0.05, one-way ANOVA with Tukey's multiple comparisons test.

Figure 2. STEMprep™-Processed Mouse Brain Microglia Are Phagocytic and Produce Cytokines upon Activation

Mouse brain tissue was processed into single-cell suspensions using STEMprep™ Mouse Brain Dissociation Kit and the STEMprep™ Tissue Dissociator, an alternative automated system, or a manual dissociation method. (A) Brain CD11b+ microglia were isolated from the single-cell suspensions using EasySep™ Mouse CD11b+ Positive Selection Kit II. (B) The isolated CD11b+ cells were incubated for 2 hours in the presence of pHrodo™ Green-conjugated E. coli BioParticles™ at 2 - 8°C (Cold) or 37°C. The fluorescence of phagocytosed BioParticles™ was measured by flow cytometry. (C) Intracellular flow cytometry staining of TNF-ɑ production by brain CD11b+ microglia cultured overnight in the presence of 3 µg/mL Brefeldin A and treated with (+) or without (-) 100 ng/mL of lipopolysaccharides (LPS). Data are presented as mean ± SD (n = 4).