Isolated hippocampal neurons in cryopreserved long-term cultures: Development of neuroarchitecture and sensitivity to NMDA

摘要: Isolated neurons in long-term culture provide a unique opportunity to address important problems neuronal development. In the present study we established conditions for cryopreservation and primary of isolated embryonic hippocampal neurons. This system was then used initial characterizations development neuroarchitecture neurotransmitter response systems. Cryoprotection with 8% dimethylsulfoxide, slow freezing, rapid thawing provided high-yield cultures which appeared normal terms cell types, mitotic ability, axonal dendritic outgrowth, sensitivity glutamate neurotoxicity. A reduced medium volume moderate elevation extracellular K+ 20 mM promoted survival through 3 weeks culture. The outgrowth axons dendrites pyramidal-like found differ over 3-week period such that continued grow at relatively constant rate while slowed during second week ceased by end 3. Developmental changes were also observed pyramidal neurotoxicity; functional kainate/quisqualate receptors first culture, responses N-methyl-D-aspartic acid (NMDA) did not appear until week. technologies reported here useful variety neuroscience.