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). The method exploits the different growth and adhesion characteristics of fibroblasts and Schwann cells under different experimental conditions such that antiproliferative (cytotoxic) drugs or time-consuming centrifugation is not needed. Standard procedures were used to obtain mixed cultures of Schwann cells, fibroblasts and neurons. After about 5 days further purification of the cells was achieved by exploiting the different responses of Schwann cells and fibroblasts to a temperature shock. Cooling the cells with cold phosphate-buffered saline (PBS), followed by pipetting cold medium directly on top of the cells ('cold jet'), resulted in specific detachment of Schwann cells and neurons, whereas fibroblasts remained securely attached. Schwann cells attached to the surface of new, uncoated culture dishes whereas neurons did not. Two cycles of the cold jet procedure resulted in nearly pure (98-100%) cultures of Schwann cells. Besides being gentle, this method is easy and fast, and because cytotoxic drugs are not used, it does not affect cell survival negatively.
Jirsová
K
K
Institute of Experimental Medicine, Academy of Sciences, Prague, Czech Republic.
Sodaar
P
P
Mandys
V
V
Bär
P R
PR
D000818:Animals; D017209:Apoptosis; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003080:Cold Temperature; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology; D005727:Ganglia, Spinal / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D012583:Schwann Cells / Q000166:cytology*