骨髓基质细胞移植促进脊髓全横断损伤区神经元轴突的再生变化  

Marrow stromal cell transplantation enhances regeneration of neuronal axon after spinal cord transection

作　　者：丁鹏[1] 薛黎萍[2] 宋晓斌[1] 李玉[1] 龙江[1] 王伟民[1] 杨智勇[1] 王进昆[1] 林荣安[3] 

机构地区：[1]昆明医学院第一附属医院神经外科,云南省昆明市650032 [2]云南省第二人民医院眼科,云南省昆明市650021 [3]新加坡国立大学解剖系,新加坡117597

出　　处：《中国组织工程研究与临床康复》2009年第49期9631-9636,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:近年来,部分学者证明骨髓基质细胞移植可促进轴突再生,改善脊髓损伤引起的运动功能障碍,但目前关于移植骨髓基质细胞如何促进轴突再生,移植细胞与再生轴突的关系尚不清楚。目的:通过免疫荧光组织化学和免疫电镜的方法,探讨移植骨髓基质细胞促进脊髓全横断损伤区轴突再生的机制。设计、时间及地点:随机对照动物实验,细胞学体内观察,于2006-03/2007-06在新加坡国立大学解剖系完成。材料:清洁级Wistar新生大鼠1只,用于骨髓基质细胞培养。清洁级成年雌性Wistar大鼠36只,无菌条件下显露、切断脊髓T10,制备脊髓全横断损伤模型。方法:通过传代法培养、纯化骨髓基质细胞。36只成年Wistar雌性大鼠随机投币法分为移植组和对照组,每组18只。移植组大鼠脊髓全横断损伤9d后以1×1011L-1的密度移植骨髓基质细胞,缺损区5μL,损伤区上、下1mm处各2.5μL,对照组动物在相同部位注射等量DMEM完全培养基,注射速度1μL/min。主要观察指标:①移植骨髓基质细胞存活、分化情况。②轴突再生情况。③移植组和对照组宿主自身的nestin、NF200、GFAP和CNP阳性细胞在脊髓损伤区存活情况。④内源性CNP阳性细胞和再生纤维关系。结果:骨髓基质细胞移植2周时,脊髓损伤区可见大量CFDA-SE标记的移植细胞,随时间延长,存活的移植细胞数目逐渐降低,考虑脊髓损伤区内大量的OX42阳性吞噬细胞/激活小胶质细胞及空洞可能影响移植细胞的存活。虽然骨髓基质细胞数目逐渐降低,骨髓基质细胞移植可促进损伤区轴突的再生,而且还可促进宿主自身的nestin、NF200、GFAP和CNP阳性细胞在脊髓损伤区存活。宿主自身CNP和许旺细胞促进损伤轴突的再生和髓鞘形成。结论:移植骨髓基质细胞移植可促进宿主自身CNP和许旺细胞在脊髓损伤区存活,后者具有促进损伤轴突再生和髓鞘形成的作用。BACKGROUND：Although previous studies have indicated that transplantation of marrow stromal cells （MSCs） has enhanced axonal regeneration and improved motor dysfunction induced by spinal cord injury.However,it is still unclear how transplanted MSCs promoted axonal regeneration and the relationship of transplanted cells and regenerated axons.OBJECTIVE：By immunofluorescence and immunoelectron microscopy,this study sought to elucidate the mechanism of promoting axonal sprouting following transplantation of MSCs into a completely transected spinal cord.DESIGN,TIME AND SETTING：The in vivo cytology randomized controlled animal experiment was performed at the Department of Anatomy of National University of Singapore from March 2006 to June 2007.MATERIALS：MSCs were isolated and purified from a Wistar neonatal rat.Model of completely transected spinal cord injury was established by transection at T10 segment with asepsis technique using 36 clean adult female Wistar rats.METHODS：MSCs were subcultured and purified.Thirty-six adult female Wistar rats were randomly divided into transplanted and control groups,with 18 animals in each group.Following 9 days of completely transected spinal cord injury,rats in the transplanted group were injected with MSCs （1×10^11/L）,5 μL in the defect region and 2.5 μL in 1 mm upper and lower the defect region.Rats in the control group were infused with an equal volume of DMEM,at the speed of 1 μL/min.MAIN OUTCOME MESSURES：Survival and differentiation of transplanted MSCs;Regeneration of axon and survival of host-derived nestin-,NF200-,GFAP-,and CNP-positive cells in control and transplanted groups;Relationship of regenerated axon and CNP-positive cells RESULTS：Two weeks after transplantation,a large number of CFDA-SE labeled MSCs were detected in the lesion site.Survival transplanted cell number was decreased over time.Abundant OX42-positive phagocytes/activated microglia and cavity might affect the survival of transplanted cells.The number of MSCs was decreased,but MS

关 键 词：脊髓损伤 骨髓基质细胞 移植 轴突 再生 

分 类 号：R394.2[医药卫生—医学遗传学]