The video demonstrates the isolation of tumor-infiltrating peripheral blood mononuclear cells from a mouse brain using mechanical dissociation and enzymatic digestion. This is followed by density gradient centrifugation to collect the immune cells for further analysis.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. Prepare density centrifugation media mix solution by placing 90 ml of 10x PBS into 264 ml of ultrapure deionized water (3.93x) in a sterile plastic container. Titrate to pH 7.0-7.2 with HCl and filter sterilize through a 0.22 µm filter. Store at 4 °C. Prepare 70% density centrifugation media by combining 18 ml of density centrifugation medi…
Representative Results
Figure 1: Isolation of Glioma-infiltrating PBMCs. (A) Strategy for the dissection of mouse brain tissue containing early-stage orthotopic glioma implants. The top panel demonstrates the sagittal bisection of the ipsilateral hemisphere away from the contralateral hemisphere. The bottom panel demonstrates the two additional coronal cuts necessary to isolate the target tissue contai…
Offenlegungen
The authors have nothing to disclose.
Materials
Dulbecco's Modification of Eagles Medium
Gibco
12430-054
With 4.5g/L D-glucose, L-glutamine, 25mM HEPES and phenol red