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bioRxiv · 10.1101/2025.08.08.666877

Protocol to isolate oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain using magnetic-activated cell sorting

Houmam, S.·Siodlak, D.·Pham, K.·Salinas, C.·Ocanas, S. R.·Freeman, W. M.·Rice, H. C.
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Abstract

The isolation of specific cell types of the brain is essential to study cell-type-specific differences in complex neurological diseases such as Alzheimers disease. This protocol isolates oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain. The process involves gentle tissue homogenization, debris removal, and sequential sorting of five distinct cell types. We validate cell purity and viability using flow cytometry and RT-qPCR. This protocol is well-suited for a range of downstream applications, including genomics, transcriptomics, and proteomics. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/666877v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1426b97org.highwire.dtl.DTLVardef@1a5bb77org.highwire.dtl.DTLVardef@1b69f32org.highwire.dtl.DTLVardef@8da416_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Houmam, S., Siodlak, D., Pham, K., Salinas, C., Ocanas, S. R., Freeman, W. M., Rice, H. C.. 2025-08-12. Protocol to isolate oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain using magnetic-activated cell sorting. https://doi.org/10.1101/2025.08.08.666877

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