神经膜细胞与人发角蛋白复合培养构建人工神经桥接体的实验研究  

Construction of artificial nerve bridge by three-dimensional culture of Schwann cells with human hair keratins

作　　者：杨俊[1] 刘巍[2] 余磊[1] 邱小忠[1] 焦培峰[1] 胡莲美[1] 武雷[1] 朴英杰[1] 钟世镇[1] 秦建强[1] 

机构地区：[1]南方医科大学解剖学研究所,广东广州510515 [2]南方医科大学病理学教研室,广东广州510515

出　　处：《第一军医大学学报》2005年第9期1128-1131,1144,共5页Journal of First Military Medical University

基　　金：广东省科技计划重大专项(2003A3020101);广东省重点实验室(2001B60107);国家自然科学基金(39970833)~~

摘　　要：目的探索神经膜(旧称许旺细胞)与人发角蛋白复合培养构建人工神经桥接体。方法用10μmol/mlBrdU标记体外培养纯化的许旺细胞与ECM凝胶修饰的人发角蛋白进行三维方向的共培养而构建成人工神经桥接体,移植入SD大鼠坐骨神经缺损处、皮下、骨骼肌中。S-100蛋白免疫组化染色鉴定许旺细胞,倒置显微镜和扫描电镜下观察许旺细胞与人发角蛋白的共培养,石蜡切片抗BrdU免疫组化染色观察体外培养纯化的许旺细胞在体内的生长情况。结果体外培养的许旺细胞能黏附于人发角蛋白上进行良好的生长,移植入神经缺损处4周后,坐骨神经缺损处、皮下、骨骼肌中的桥接体内的人发角蛋白开始降解,黏附于该人发角蛋白上的许旺细胞均见成活并分裂增殖。结论许旺细胞可与人发角蛋白进行三维培养,构建成人工神经桥接体,填补神经缺损。Objective To culture Schwann cells （SCs） and human hair keratins （HHKs） for artificial nerve bridge construction. Methods SCs were purified by primary culture and labeled with BrdU, which were then cultured with HHKs decorated by ECM. The artificial nerve bridge was implanted into the defect of sciatic nerve, beneath the skin, and in the skeletal muscles of SD rat, respectively. The morphology of the SCs cultured with HHKs was monitored by inverted microscope and evaluated by immunocytochemical staining. Growth of BrdU-labeled SCs in vivo was observed by immunocytochemical staining on paraffin sections. Results In vitro cultured SCs were capable of adhering to HHKs and grew well four weeks after implantation. The HHK component in the artificial nerve bridge underwent degradation in the defect of the sciatic nerve, beneath the skin, and in the skeletal muscles of SD rat, and SC survival and proliferation were verified. Conclusion SCs can survive in three-dimensional culture with HHKs for construction of artificial nerve bridge to repair nerve defects.

关 键 词：神经膜细胞 人发角蛋白 周围神经 生物工程 

分 类 号：R318.0[医药卫生—生物医学工程]