关键词: 乙二醇-双-(2-氨基乙醚)四乙酸；SD成年大鼠；坐骨神经；许旺细胞；纯化

Abstract:

 Objective  To investigate the efficiency of ethylene diamine tetraacetic acid (EDTA) to purify SD Schwann cells. Methods  Firstly, epineurium and perineurium of SD bilateral sciatic nerves were peeled off under high power microscope. Afterwards the nerves without epineurium and perineurium were digested using collagenase NB4 for 30min. Then the supernatant was discarded after centrifuge at 600g. The resuspended sciatic verve fragments were cultured in medium supplemented with nerve growth factor for 1 week to get pre-degenerated tissues. The primary Schwann cells were harvested after the digestion of pre-degenerated sciatic nerves tissues with trypsin. Sub-confluent cells were treated by EDTA-PBS for 3 minutes and observed under microscope. Then, the cell pellet was resuspended and cultured in Schwann cell medium for 2 days. The purity of the subconfluent Schwann cells treated with EDTA-PBS was demonstrated by immunofluorescence staining with P75. Results  The purity of Schwann cells was up to 99% or so after twice purification by EDTA. Conclusion  EDTA is an efficient and effective method to purify SD Schwann cells.

Key words: Ethylene glycol-double-(2-aminoethyl ether) tetra-acetic acid; Adult SD rats; Sciatic nerves;Schwann cells; Purification