鼻粘膜干细胞来源施万样细胞与PC12高分化细胞共培养研究  

The co-culture research of the Schwann-like cells differentiated from nasal mucosa stem cells with high differentiated PC12 cells

作　　者：张健[1] 高鑫[1] 张志坚[2] 邹红军[1] 刘锦波[1] 

机构地区：[1]苏州大学附属第三医院骨科,常州213003 [2]江苏大学基础医学与医学技术学院,镇江212013

出　　处：《中国矫形外科杂志》2014年第10期911-917,共7页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:30570981和30571878);常州科技局基金资助项目(编号:CJ20122027)

摘　　要：[目的]研究大鼠鼻粘膜来源干细胞(REMSCs)分化为施万样细胞与PC12高分化细胞共培养后髓鞘形成及轴突生长情况。[方法]取大鼠鼻甲组织分离、培养及鉴定REMSCs,使用Heregulin、血小板源性生长因子、成纤维细胞生长因子和Forskolin将REMSCs诱导分化为施万样细胞。将分化后形成的施万样细胞与PC12高分化细胞共培养。倒置显微镜下观察形态,免疫细胞化学染色检测细胞表面标志,电镜观察髓鞘形成能力,Western blot检测MBP。[结果]大鼠REMSCs表达外胚层标志物:nestin,SOX10和Vimentin。REMSCs诱导后形成的施万细胞成双极纺锤状,并且能够在体外持续增殖。倒置显微镜观察到施万样细胞沿PC12高分化细胞轴突生长并且促进PC12高分化细胞轴突延长,电镜显示施万样细胞和PC12高分化细胞形成髓鞘样结构。Western blot显示MBP在共培养3,6,9 d呈递增趋势。[结论]从下鼻甲分离的REMSCs诱导后可形成施万样细胞,施万样细胞具有形成髓鞘及促进轴突生长能力。[ Objective] To investigate the ability of myelin formation and promoting axon outgrowth of Schwann-like cells（SC-like ceils） derived from nasal mucosa stem cells by co-culturing SC-like cells with high differentiated PC12 cells. [ Method] Rat REMSCs were harvested, cultured and identified from inferior nasal concha. Heregulin, basic fibroblast growth factor,platelet-derived growth factor and forskolin were used to induce REMSCs into SC-like cells. Then SC-like cells were co-culturedwith high differentiated PC12 cells. The morphology of SC-like cells was observed by inverted microscope. Immunofluorescencewere used to detect the surface markers of SC-tike cells and the axon of high differentiated PC12 cells. Transmission electron mi-croscope was used to observe myelin structure of SC-like cells. Western blot was used to detected the myelin basic protein（MBP）. [ Result ] REMSCs expressed ectoderm markers ：nestin, SOX10 and Vimnetin. SC-like cells differentiated from REMSCsexhibited a bi-polar, spindle-like morphology and could proliferate in vitro. Inverted microscope showed that SC-like cells grew a-long the axon of high differentiated PC12 cells and promoted the outhgrowth of the axon. Transmission electron microscopeshowed that SC-like cells could form myelin structure with the axon of high differentiated PC12 cells. Western blot showed thatMBP was in increasing tend when detecting the level of MBP at 3,6,9 days. [ Conclusion] REMSCs derived from inferior nasalconcha could differentiate into SC-like cells. SC-like cells could form myelin and promote axon growth in vitro.

关 键 词：鼻粘膜干细胞 施万样细胞 髓鞘 高分化PC12细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]