HBSS w/ or w/o calcium for dissecting cortical neurons? - (Jul/23/2009 )
I am interested in culturing primary cortical neurons from E18. Somebody advised me to triturate the cortices in HBSS with calcium, magnesium, but I've noticed that a lot of people use calcium, magnesium-free HBSS instead. Would using one over the other as a dissecting medium affect cell viability, culture conditions? (The cultures will eventually be maintained in Neurobasal with B27.)
Calcium & magnesium ions are needed for cell-cell and cell-matrix adhesion. If you would really want to dissociate cells, it's best to use Ca2+ & Mg2+ free buffer. For the second Q: Would using one over the other as a dissecting medium affect cell viability, culture conditions? I can't really answer this with confident; however my take would be that it wouldn't effect a difference.