Bulk separation of neurones and glia: A comparison of techniques

Abstract

Three different techniques of preparation of neurones and neuropil (glia) from rat cortex, those of Rose, Norton and Poduslo, and Sellinger and Azcurra, were compared according to the proposed criteria of purity, integrity and yield. Morphologically the cells prepared by the Norton and Poduslo and Sellinger and Azcurra methods appeared better and the fractions were purer than Rose method I, though not Rose method II. Based on carbonic anhydrase ratios, the glial contamination of neurones was 11% in Rose cells, 24.8% in Norton and Poduslo cells and 8.3% in Sellinger and Azcurra cells. Lactate dehydrogenase activity was 25% in Sellinger and Azcurra cell suspension and 37% in Norton and Poduslo cell suspension, of that in the Rose suspension. Oxygen consumption and carbon dioxide production in Sellinger and Azcurra suspension was only 15% and in Norton and Poduslo cell suspension 50%, of that in the Rose technique. The metabolic damage observed in the Sellinger and Azcurra preparation could be partially averted by alteration of the disaggregation medium. The relative utility of the techniques is discussed.