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Characterization of neuronal and glial fractions separated in sucrose and ficoll media.

Rat brain cerebral cortices were dispersed by a collagenase-hyaluronidase treatment and the suspensions obtained were fractionated in sucrose and Ficoll systems by centrifugation. For sucrose a short sedimentation run was followed by a density gradient; for Ficoll only a density gradient was used. Isolated cell populations were examined by light microscopy and differential cell counts were taken. Separation in sucrose yielded more homogenous but smaller fractions. In Ficoll up to 37 million neuronal and glial cells could be collected from 5 cortices. Cells isolated in sucrose and Ficoll contained 70 and some 50 pg protein, respectively. 
 
 
 
Neuronal and glial cells separated in sucrose and Ficoll systems accumulated potassium, and were metabolically active. Both respiration and incorporation of radioactive leucine into protein were clearly more active in neuronal fractions. In Krebs-Ringer phosphate buffer containing 10 mM glucose, neurons respired 946 and 875, and glia 706 and 396 nmoles oxygen/mg protlhr after sucrose and Ficoll separation respectively. Incorporation of radioactive leucine into protein was 5370 and 4980 dpm/mg prot for neurons, and 3110 and 4480 dpm/mg prot for glia. On the other hand, no clear difference was found in TCA-soluble radioactivities be tween neuronal and glial fractions.