Abstract:

BACKGROUND: Peripheral nerve tissue engineering needs a large number of Schwann cells. In previous studies, lack of normal human nervous tissue, so animal (rat, rabbit, et al) nervous tissues are commonly used to isolate Schwann cells, but as xeno-cells it is limited in clinical application.

OBJECTIVE: To investigate an effective technique for isolation, cultivation and purification of human Schwann cells of normal peripheral nerves cultured in vitro.

METHODS: Normal peripheral nerves were obtained from the surgery of cerebral palsy patients. Schwann cells were cultured with enzymatic digestion culture method and differential attachment method. Tissues were cut into pieces and incubated in medium supplemented with fetal bovine serum, collagenase and Dispase enzyme, centrifuged. Tissue blocks were placed in the medium, triturated into monoplast suspension, and then moved into a DMEM Petri dish containing polylysine, supplemented with basic fibroblast growth factor. When adherent cells were confluent about 85%-90%, cells could subculture. Schwann cells were counted by Trypan Blue coloring method at 2, 3, 4, 5, 6, 7, 8, 9, 10. The purity of Schwann cells was identified through S-100 protein immunohistochemistry staining. 

RESULTS AND CONCLUSION: More than 0.5×10⁸/L Schwann cells were detected after four days under a microscope. Following the third passage, the number of Schwann cells was over 9×10⁸/L. The purity of Schwann cell population was up to 85%. Results suggested that plenty and purified human Schwann cells could be obtained by enzymatic digestion culture and differential attachment methods in a short time, which can be used for the source of peripheral nerve tissue engineering.