The use of an artificial nerve conduit containing viable Schwann cells (SCs) is one of the most promising approaches to repair nerve injuries. Obtaining a large number of viable SCs in a short period is demanded for the clinical use of this technique. However, the previous methods using mitogens are not clinically acceptable, and other methods that do not require mitogens, failed to isolate adult SCs effectively or required a long period of time. In this study, we have developed a novel technique to isolate SCs from adult rat peripheral nerves for an artificial nerve conduit without mitogens, which has produced a total number of 1.21×105 cells per mg, with an average purity of 93.0±0.58% at 21 days in vitro. The Bottenstein-Sato (BS) medium used in this study, had originally been developed for oligodendrocyte culture, but here it is shown to have an effect on SC proliferation and survival. By changing fetal bovine serum (FBS) concentrations from 0 to 10% serially, SCs could be isolated maximally from the predegenerated nerves while suppressing fibroblast overgrowth. The combination of this technique and the altered medium promoted the migration and proliferation of SCs selectively by utilizing the supporting cells of SCs instead of discarding them by changing the culture dishes and media.
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