A modified procedure for isolation of astrocyte- and neuron-enriched fractions from rat brain.

Abstract

AbstractOur previously published method for isolation of neurons with extensive processes (Farooqet al., 1977) has been modified to permit the isolation of both astrocyte‐ and neuron‐enriched fractions. Rat cerebral tissue is incubated with acetylated trypsin and disrupted. The cell suspension is separated first by differential centrifugation and then by gradient centrifugation on discontinuous Ficoll gradients. The method is reproducible and is applicable equally well to immature and adult animals. The yield of astrocytes of 57% particle purity, and higher weight purity, is 4–7 × 106 cells/brain, amounting to 1.5–2.0 mg of protein. The astrocytes appear to be a mixture of fibrous and protoplasmic types. The yield of neurons of 90% particle purity is 10–14 × 106 cells/brain, amounting to 2.4–3.0 mg of protein. A total yield of neurons of 28–37 × 106 cells/brain can be obtained at 70% purity. These preparations have been characterized by light microscopy and protein, RNA and DNA content.