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A simplified procedure for isolation of primary murine microglia

There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a s...

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Detalles Bibliográficos
Autores principales: Scott, Nathan, Witt, Ken, Schober, Joseph M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9761776/
https://www.ncbi.nlm.nih.gov/pubmed/36398847
http://dx.doi.org/10.2144/btn-2022-0054
Descripción
Sumario:There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a single step to isolate the microglia from the mixed cell population. Once the microglia were isolated, we characterized cell purity, microglial morphology and phagocytic activity. The single-step protocol, without the need for additional astrocyte or oligodendrocyte separation, allows microglial cells to be used immediately for experimental purposes. The protocol is low-cost and can be performed in any lab with standard cell-culture equipment.