Abstract:

BACKGROUND: Tissue engineering methods to construct nerve bridge graft for repairing nerve injury requires massive purified Schwann cells cultured in vitro.
OBJECTIVE: To compare the purity and quality of Schwann cells obtained with the pre-injury method and improved passage method.
METHODS: (1) Pre-injury method: Sciatic nerve of Sprague-Dawley neonatal rats was pre-injured for 3 days and removed, the epineurium was isolated and digested with trypsin and collagenase. Schwann cells were cultured in the culture medium after fibroblasts were eliminated with differential adhesion method. (2) Improved passage method: Sciatic nerve was directly removed from Sprague-Dawley neonatal rats and the epineurium was isolated. Double-enzyme digestion and single-enzyme digestion methods were adopted to primarily culture Schwann cells for 5-7 days. Schwann cells were purified with a single enzyme rapid digestion and centrifugation.
RESULTS AND CONCLUSION: Both pre-injury and improved passage methods obtained 95% purity of Schwann cells cultured in vitro, with no significant difference (P > 0.05). The resultant Schwann cells obtained by two methods showed normal morphology, high quantity, high purity and productive growth. These experimental findings imply that both pre-injury and improved passage methods are ideal approaches for in vitro culture of Schwann cells with high quality and high purity.