Mononuclear Cell Isolation from Mouse Brain: A Density Gradient Centrifugation Technique to Isolate Brain Resident Mononuclear Cells
Published: April 30, 2023
Abstract
Source: Ji, Z. et al. Flow Cytometric Analysis of Lymphocyte Infiltration in Central Nervous System during Experimental Autoimmune Encephalomyelitis. J. Vis. Exp. (2020)
In this video, we perform the isolation of mononuclear cells from a mouse’s brain using density gradient centrifugation. The isolated mononuclear cell suspension can be used for further downstream analysis.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Single-Cell Suspension Preparation from Brain Dilute density gradient medium in 9:1 ratio with PBS in a 15 mL conical tube to yield a final 100% solution. Anesthetize mice at the peak of EAE with an intraperitoneal injection of 1% sodium pentobarbital (50 mg/kg) and perfuse intracardially with 20 mL of sterile…
Representative Results
Figure 1: Schematic of the Percoll gradient setup for isolation of mononuclear cells.
開示
The authors have nothing to disclose.
