Rapid and robust isolation of microglia and vascular cells from brain subregions for integrative single-cell analyses.
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Preka E, Lastra Romero A, Sun Y, Onetti Vilalta Y, Seitz T, Fragkopoulou A, Betsholtz C, Osman AM, Blomgren K
Heliyon 10 (16) e35838 [2024-08-30; online 2024-08-05]
Cell isolation protocols from brain tissue include prolonged ex vivo processing durations, rendering them suboptimal for transcriptomic studies. Particularly for microglia and vascular cells, current isolation methods produce lower yields, necessitating addition of an enrichment step, and use of large tissue volumes - in most cases whole brain tissue - to obtain sufficient yields. Here, we developed a simple, rapid, and reproducible cell isolation method for generating single-cell suspensions from micro-dissected brain regions, enriched for microglia and vascular cells, without an enrichment step. Cells isolated using this method are suitable for molecular profiling studies using 10 × Genomics Chromium single-cell RNA sequencing with high reproducibility. Our method is valuable for longitudinal unbiased molecular profiling of microglia and vascular cells within different brain regions, spanning multiple time points across physiological development or disease progression.
Bioinformatics Support for Computational Resources [Service]
NGI Stockholm (Genomics Applications) [Service]
NGI Stockholm (Genomics Production) [Service]
National Genomics Infrastructure [Service]
PubMed 39211933
DOI 10.1016/j.heliyon.2024.e35838
Crossref 10.1016/j.heliyon.2024.e35838
pmc: PMC11357767
pii: S2405-8440(24)11869-5