Novel method for culturing Schwann cells from adult mouse sciatic nerve in vitro

Abstract

Schwann cells (SCs) are important in the recovery of peripheral nerve injury and are valuable cells for the tissue engineering of artificial neurons. Clinical applications that require pure SCs in large quantities are limited since human and mouse SCs do not attach well to the wall of the culture dish and have low proliferative potential. To obtain high quantities of highly pure SCs, we developed a new method for culturing SCs from the mouse sciatic nerve in vitro. Approximately 1.5 cm of the bilateral sciatic nerve of a c57 adult mouse was surgically removed and pre-cultured in DMEM containing either 10% FBS or growth factors. One week later, the in vitro SC culture was observed using light microscopy following enzyme digestion. Cell numbers and cell attachment were examined. The purity of the SCs was determined using s100β and p75NTR staining. Sciatic nerves that had not been pre-cultured were used as the control group. When the excised tissue was pre-cultured in vitro, high yields of SCs were obtained. The SCs were more likely to adhere and the purity was approximately 98% at the p1 generation following simple purification steps, which was significantly higher than the purity obtained from the control group. The pre-culturing of the sciatic nerve prior to in vitro tissue culturing significantly increased the quantity and quality of the SCs.