An alternative method for obtaining high-viability cell suspensions from neonatal mouse brain.
Cultured brain cells have contributed greatly to our understanding of a variety of neurobiological processes. The ability to culture brain tissue is important for studying cellular processes underlying unique neural properties. Traditional culturing techniques commonly involve triturating tissue through glass Pastuer pipets, which are inappropriate for use with potentially biohazardous materials. We therefore developed an alternative method for dissociating brain tissue. The protocol combines enzymatic digestion and mechanical dissociation with additives to the dissection medium that protect the cells against other sources of injury, including glutamate neurotoxicity, oxidative damage, and excessively alkaline pH. We find this method works well with post-natal mouse brain, consistently giving cell viabilities in the range of 92-99% and an average yield of 3.1 x 10(6) cells per mouse.
Martin
F C
FC
Department of Pathology, University of California at San Diego, La Jolla, CA 92093-0612.
Wiley
C A
CA
D000818:Animals; D000831:Animals, Newborn / Q000033:anatomy & histology*; D001921:Brain / Q000166:cytology* / Q000254:growth & development; D002478:Cells, Cultured; D003584:Cytological Techniques; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling