Experimental study of separation and purification of Schwann cells by immunomagnetic heads method / 中国矫形外科杂志

ABSTRACT

[Objective]To introduce a method to obtain Schwann cells from newly born SD rats massively and purely by immunomagnetic heads method.[Method]SD rats that had been born within 5 to 7 days were used. Their bilateral sciatic nerves were dissected under sterile condition. Under 16?microscope the nerve fascicles and the epineurium were carefully extracted in oder to get the nerve tract without impurity cell. Then the nerve tract was cut into small particle about 1 mm3.Two enzymes (0.25% trypsinase and 0.2% collagenaseⅠ)were used to digest the particle of sciatic nerve specimens twice. After using 20% fetal bovine serum to stop the process of digestion,centrifugate(1000r/min,5 min) and DF12 culture medium was added into the precipitation.Seven days later,Schwann cells were purified with immunomagnetic heads.During the whole process,the change of Schwann cells was observed under the inverted biological microscop and vital force of Schwann cells was evaluated.The growth curve of Schwann cell was drawn with MTT.The Schwann cells was identicated under immunofluorescent test and record the purity.[Result]The cell separated from the SD rat's sciatic nerve and cultured in incubator was affirmed as Schwann cells.Immunomagnetic heads can be used to purify the Schwann cells.The 96% vigor and 98% pure Schwann cell cultures were generated and passaged after two days.[Conclusion]This method can obtain massive purified normal schwann cells to satisfy the need of tissue-engineered bioartificial nerve graft.