Simultaneous Flow Cytometric Characterization of Multiple Cell Types Retrieved from Mouse Brain/Spinal Cord Through...

Area of Science:

• Neuroscience
• Immunology
• Cell Biology

Background:

• Viral vectors and nanomaterials offer novel ways to study the central nervous system (CNS).
• Optimizing these tools requires methods to assess CNS and cell-specific targeting.
• Current methods may not efficiently quantify cell subtypes after administration.

Purpose of the Study:

• To present a high-throughput flow cytometry protocol for quantifying CNS cell subtypes.
• To compare tissue homogenization methods for cell yield, viability, and composition.
• To provide a tool for optimizing viral vector and nanoparticle delivery in the CNS.

Main Methods:

• Isolation of single-cell suspensions from mouse brain and spinal cord.
• Utilizing flow cytometry for high-throughput, multiplexed cell quantification.
• Analysis of microglia/macrophages, lymphocytes, astrocytes, oligodendrocytes, neurons, and endothelial cells.

Main Results:

• The protocol enables straightforward quantification of diverse CNS cell subtypes.
• Differences in cell yield, viability, and composition were observed between two homogenization methods.
• The choice of homogenization method impacts cell recovery and suitability for downstream applications.

Conclusions:

• The developed flow cytometry protocol is valuable for neuroscientists studying CNS targeting.
• It facilitates the selection of optimal tissue processing methods based on cell type and application.
• This method supports cell sorting for further analyses like gene expression and functional assays in neurodegenerative diseases, pharmacology, and gene therapy.