Abstract:Objective: To investigate the effect of human embryonic stem cell-derived neurospheres conditioned medium(hESCN-CM) on the proliferation and migration of Schwann cells and its potential mechanisms. Methods: The bilateral sciatic nerves of 3-5 days old SD rats were isolated for Schwann cells, which were identified by immunofluorescence. Schwann cells were stimulated with 0%, 50% and 100% hESCN-CM and the effect of hESCN-CM on the proliferation of Schwann cells was measured by CCK-8 assay. The migration of Schwann cells was measured by would healing assay. The mRNA expression of Nt3, Vegf, Ngf, Bfgf and Igf-1 was determined by quantitative real-time PCR(qRT-PCR). Results: Immunofluorescence staining showed that specific marker S100 expression was detected in 99% of all cells. The CCK-8 assay showed that 50% hESCN-CM promoted the proliferation of Schwann cells and inhibited proliferation at 100% concentration. Wound healing assay suggested that 50% hESCN-CM and 100% hESCN-CM promoted migration in condition of culturing for 6 h and 50% hESCN-CM promoted migration at 12 h; while showed no significance between 0% hESCN-CM and 100% hESCN-CM at 12 hours. qRT-PCR demonstrated that compared with 0% hESCN-CM, 50% hESCN-CM showed higher mRNA expression levels of Nt3, Vegf, Ngf, Bfgf. Conclusion: hESCN-CM could promote the proliferation and migration of rat Schwann cells, which may be attributed to up-regulation of mRNA expression of Nt3, Vegf, Ngf and Bfgf.
[Key words]conditioned medium; proliferation; migration; neurospheres; Schwann cells
收稿日期: 2018-12-09
基金资助:
国家自然科学基金资助项目(81571221)
通讯作者:
胡嘉波(通讯作者),教授,硕士生导师,E-mail: hu@ujs.edu.cn
作者简介: 纪贤严(1988—),女,硕士研究生;
引用本文:
纪贤严, 高建一, 叶开, 张磊, 曹江素, 胡嘉波. 人胚胎干细胞源神经球条件培养液对大鼠施万细胞增殖和迁移的影响[J]. 江苏大学学报:医学版, 2019, 29(03): 189-194.
JI Xian-Yan, Gao-Jian-Yi, Ye-Kai, Zhang-Lei, Cao-Jiang-Su, Hu-Jia-Bo. Effect of human embryonic stem cell-derived neurospheres conditioned medium on the proliferation and migration of rat Schwann cells. Journal of Jiangsu University(Medicine Edition), 2019, 29(03): 189-194.