关键词: 许旺细胞, 冷酶消化法, 分离, 细胞培养, 免疫组织化学

Abstract:

BACKGROUND: Currently, conventional enzymatic digestion methods are commonly used to isolate and culture Schwann cells. Because trypsinase has toxic effects on Schwann cells in 30 minutes at 37 ℃, the amount and purity of Schwann cells can have a certain impact.
OBJECTIVE: To improve a novel method to rapidly isolate and culture a great amount of Schwann cells.
METHODS: Schwann cells were isolated from newborn rat sciatic nerve digested with trypsinase at 4 ℃, 6 to 18 hours, and then cultured for some time. MTT was used to draw cell growth curve. Hematoxylin-eosin staining and immunohistochemistry were used to observe cell morphology, and positive cells were counted.
RESULTS AND CONCLUSION: The quantity of Schwann cells was over 1×106 and the purity of Schwann cells was over 90% assessed by anti-s100 protein immunohistochemistry stainer. From the morphology of cultured cells and cells growth curve, the cold enzyme digestion method was simply and effective, the purified cells had good biologic characteristics. In conclusion, the novel method can satisfy the clinical need such as isolating and culturing time, the quantity and purity of cells and the bioactivity of Schwann cells.