Purification of newborn rat Schwann cells by multiple digestions with low-concentration trypsin for differential cell detachment

Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies. Key words: Sehwann cells; Purification; Collagenase digestion of differential detachment