In vitro predegeneration of peripheral nerve; the effect of predegeneration period on rat Schwann cell cultures.

Abstract

Peripheral nerve predegeneration has been used as a tool to improve the in vitro cultivation of Schwann cells. The process of predegeneration may be accomplished either in vivo or in vitro. In previously published studies, various predegeneration periods were used, ranging from a few days until up to 5 weeks. The present study systematically evaluated the effect of various durations of in vitro predegeneration on the efficacy of Schwann cell cultivation. The sciatic nerves of adult Wistar rats were harvested and the explanted nerve pieces were maintained in the predegeneration medium for different predegeneration periods. In group A, the dissociation was performed immediately after harvesting. In groups B, C and D, the predegeneration periods were 2, 4 and 6 weeks, respectively. During the predegeneration period, the tissue pieces were repeatedly transferred into new dishes. Afterwards, the nerve tissue was enzymatically dissociated and the cells were seeded onto a six-well culture plate at a defined density. After 3–4 days of incubation, the cultures were passaged by means of the cold jet technique and the cell cultivation was continued for another 21 days. It was observed that the cell cultures in groups A and B were rapidly overgrown by fibroblasts. In group C, numerous wells contained a highly enriched Schwann cell population that had formed a typical monolayer, but in a fraction of the dishes, cultures were debased by fibroblast overgrowth. In group D, all of the cultures had enriched Schwann cell populations. In the experiments of the present study, the positive effect of predegeneration was observed only when the predegeneration periods lasted for 4 weeks or longer. It was concluded that the longer predegeneration periods activated Schwann cells and/or depleted the fibroblast proliferation capacity.