A Novel Approach to Rapid and High-efficiency Purification of Schwann Cells in Rats

Objective To explore a simple method for high-purity enrichment of Schwann cells (SCs). Methods The sciatic nerve was dissected from both limbs of the neonatal Sprague-Dawley (SD) rat (3-5 days old). The nerve was first digested by dispase Ⅱ to remove epineurium and perineurium, and then by collagenase NB4 to obtain primary SCs. The cells were centrifuged and cultured until confluence in a flask. Then the flask was placed in refrigerator (4 ℃) for 5 minutes. At such a low temperature, SCs mostly detached from the inner wall, while the other cells remained adherent. The SC purity was identified by S100 immunofluorescence double staining and flow cytometry. Results The purity was above 85% after enzyme digestion, and reached 99% after low-temperature treatment. Conclusion Enzyme digestion in conjunction with low temperature treatment is a simple and efficient way to obtain Schwann cells with high purity.