Isolating Mouse Brain Vessels: A Protocol to Isolate Intact Vessels from Murine Brain Sample

Abstract

Source: Boulay, A. C. et al. Purification of Mouse Brain Vessels. J. Vis. Exp. (2015)

This video demonstrates the isolation of intact vessels from a mouse's brain that constitute the blood-brain barrier (BBB) along with neurons and glial cells. The isolated vessels can be used as an in vitro model to study their role in maintaining brain homeostasis.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Dissection NOTE: Sterile conditions are not required unless vessels are used for cell culture purposes. Prepare isolation vessel solutions: B1, add 1.5 mL of HEPES 1M to 150 mL of HBSS; B2, add 3.6 g of Dextran to 20 mL of B1; B3, add 1 g of BSA to 100 mL of B1. …

Materials

Tissue Grinder Size C	Thomas scientific	3431E25
Centrifuge 5415 R Eppendorf
Centrifuge 5810 R Eppendorf 5811000320
High-performance, Modular Stereomicroscope	Leica MZ6
Low binding tips (P1000) Sorenson BioScience 14200T
Swinnex 47 mm filter holder PP 8/Pk	Millipore	SX0004700
Nylon net filter disc Hydrophilic 20 μm 47 mm 100/Pk	Millipore	NY2004700
Nylon net filter disc Hydrophilic 100 μm 47 mm 100/Pk	Millipore	NY1H04700
PARAFILM M (roll size 4 in. × 125 ft)	Sigma	P7793-1EA
HBSS, no calcium, no magnesium, no phenol red	Life technology	14175-129
HEPES (1M)	Life technology	15630056
Dextran from Leuconostoc spp. Mr ~70,000	Sigma	31390
Bovine serum albumin	Sigma	A2153
10x PBS	Euromedex	ET330