COLD JET - A METHOD TO OBTAIN PURE SCHWANN-CELL CULTURES WITHOUT THE NEED FOR CYTOTOXIC, APOPTOSIS-INDUCING DRUG-TREATMENT

This paper presents a new and gentle method to separate Schwann cells from fibroblasts obtained from foetal rat dorsal root ganglia (DRG). T he method exploits the different growth and adhesion characteristics o f fibroblasts and Schwann cells under different experimental condition s such that antiproliferative (cytotoxic) drugs or time-consuming cent rifugation is not needed. Standard procedures were used to obtain mixe d cultures of Schwann cells, fibroblasts and neurons. After about 5 da ys further purification of the cells was achieved by exploiting the di fferent responses of Schwann cells and fibroblasts to a temperature sh ock. Cooling the cells with cold phosphate-buffered saline (PBS), foll owed by pipetting cold medium directly on top of the cells ('cold jet' ), resulted in specific detachment of Schwann cells and neurons, where as fibroblasts remained securely attached. Schwann cells attached to t he surface of new, uncoated culture dishes whereas neurons did not. Tw o cycles of the cold jet procedure resulted in nearly pure (98-100%) c ultures of Schwann cells. Besides being gentle, this method is easy an d fast, and because cytotoxic drugs are not used, it does not affect c ell survival negatively. (C) 1997 Elsevier Science B.V.