Comparative study on different culture methods to get stable primary adult Schwann cells

Abstract

Objective To seek a stable,rapid and effective method in order to obtain large number of pure adult Schwann cells (Scs) in vitro. Methods Bilateral sciatic nerves of each adult SD rat were crushed several times to establish the degeneration animal model.One week later.the degenerated nerves were detected and prepared to collagenase/dispase double enzyme digestion method,trypsin/collagenase double enzyme digestion method,single enzyme digestion for half-piece method,respectively.The S-100 immunocytochemistry was used to detect the purity of Scs. Results The confluence of cells was happened within 3-5 days in collagenase/dispase double enzyme digestion method.At time of confluence.about 70%-80%cells showed S-100 positive in this method.The quantities of cells from the degenerative nerve pieces cuhured hy single enzyme digestion were not uniform within 3-5 days.cells were confluent within 10-12 d anti less than 40%cells showed S-100 positive at time of confluence.Most of tissue pieces were tangled and cells were lost in trypsin/collagenase double enzyme digestion method.Only 3-5 cells in every 100 fiehts couht smwive in vitro,confluence is diffieult.Conclusion The collagenase/dispase double enzvme digestion method was the most stable and effective method to obtain large number of pure adult Schwann cells in vitro rapidly among the present three methods. Key words: Nerve degeneration; Schwann cells; Cell culture; Rat