过表达Shootin1的骨髓间充质干细胞移植治疗脊髓损伤.doc.doc

过表达Shootin1的骨髓间充质干细胞移植治疗脊髓损伤 张 伟1，朱肖奇1，张德才2(1南方医科大学附属深圳宝安医院，广东省深圳市 518101；2深圳市罗湖区中医院，广东省深圳市518001) 引用本文：张伟，朱肖奇，张德才. 过表达Shootin1的骨髓间充质干细胞移植治疗脊髓损伤[J].中国组织工程研究，2016，20(50):7507-7517. DOI: 10.3969/j.issn.2095-4344.2016.50.009 ORCID: 0000-0001-9004-9374(朱肖奇) 文章快速阅读： 文题释义： 慢病毒：是指以人类免疫缺陷病毒1(HIV-1)来源的一种病毒载体，慢病毒载体包含了包装、转染、稳定整合所需要的遗传信息，是慢病毒载体系统的主要组成部分。携带有外源基因的慢病毒载体在慢病毒包装质粒、细胞系的辅助下，经过病毒包装成为有感染力的病毒颗粒，通过感染细胞或活体组织，实现外源基因在细胞或活体组织中表达。 转染：是指真核细胞由于外源DNA掺入而获得新的遗传标志的过程。常规转染技术可分为两大类，一类是瞬时转染，指外源DNA/RNA不整合到宿主染色体中，因此一个宿主细胞中可存在多个拷贝数，产生高水平的表达，但通常只持续几天，多用于启动子和其他调控元件的分析。一类是稳定转染(永久转染)，指外源DNA既可以整合到宿主染色体中，也可能作为一种游离体存在。线性DNA比超螺旋DNA转入量低但整合率高。外源DNA整合到染色体中概率很小，大约1/104转染细胞能整合，通常需要一些选择性标记，得到稳定转染的同源细胞系。 Transplantation of bone marrow mesenchymal stem cells overexpressing Shootin1 for treatment of spinal cord injury Zhang Wei1, Zhu Xiao-qi1, Zhang De-cai2 (1Affiliated Baoan Hospital of Shenzhen, Southern Medical University, Shenzhen 518101, Guangdong Province, China; 2Luohu District Hospital of Traditional Chinese Medicine, Shenzhen 518001, Guangdong Province, China) Abstract BACKGROUND: Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cells (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cell transplantation. OBJECTIVE: To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS: BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cells under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Allen’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the