Isolation and Purification of Schwann Cells from Human Peripheral Nerves by Cold Jet Technique

Abstract

Schwann cells (SCs) play an important role in peripheral nerve regeneration. However, established methods for SC preparation have limitations in that they are non-specific and time-consuming, as well as having a low yield and high cost. The purpose of this study was to assess the amount and purity of SCs isolated from human peripheral nerve tissue using the cold jet technique. The cold jet technique is an SC isolation and purification method that exploits the different responses of the SCs and fibroblasts to temperature shock. Specifically, the temperature shock causes specific detachment of SCs from fibroblasts. A total of 32 human peripheral nerve tissues (sensory and motor) were obtained from reconstruction procedures for oral cancer. After the length and weight of nerve segments were measured, the nerve fascicles were digested and SCs were isolated and purified by the cold jet technique. The purity of SCs was confirmed by morphology, reverse-transcription polymerase chain reaction, immunohistochemistry and flow cytometry. Phase-contrast photomicrographs, S100 mRNA expression rates and immunohistochemical staining show that the SC purification rate was greater with the cold jet technique. Flow cytometry showed the percentage (%) gated value increased from 56.10 to 95.95%. On average, 2.48±2.57 x 106 SC/g or 0.16±0.18 x 106 SC/cm were harvested efficiently from human peripheral nerve segments, where the SC yields of sensory and motor nerves did not show a significant difference with the cold jet technique. Also, there was no difference in SC yields depending on gender. The cold jet technique was an efficient method for SC isolation and purification from the human peripheral nerves. The determined amount and the level of purity of the SC harvested by this method may be helpful for tissue engineering or cell therapy.