Abstract: The free-serum culture method was presented to remove fibroblast in nerve tissue. Sciatic nerves and olfactory bulbs from 2 d old rats were dissected, treated with trypsin and collagenase, followed by centrifugation and cultured in DMEM with 10% foetal calf serum. If fibroblast rapidly multiplied to form a confluent monolayer, the media was changed into free-serum DMEM. The process was done again. S-100 immunofluorescent staining was used to identify the purity of Schwann cells and olfactory ensheath cells. Results showed that fibroblast rapidly died in free-serum media, the purity of Schwann cells and olfactory ensheath cells were up to 98%～99% using immunofluorescent staining. The findings prove that it is a good method to remove fibroblast in nerve tissue culture because the process is simple and is operated easily.

Key words: fibroblast, free-serum culture, immunofluorescence staining