不同数量BMSCs对大鼠背根神经节生长影响的实验研究  

EFFECT OF DIFFERENT NUMBER OF BONE MARROW MESENCHYMAL STEM CELLS ON GROWTH OF RAT DORSAL ROOT GANGLIA IN VITRO

作　　者：许文静[1] 赵喆[2] 赵斌[1] 王玉[1] 彭江[1] 张莉[1] 陈继凤[1] 卢世璧[1] 

机构地区：[1]解放军总医院全军骨科研究所,北京100853 [2]武警总医院骨科

出　　处：《中国修复重建外科杂志》2011年第10期1245-1249,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划(863)资助项目(2009AA03Z312);国家科技支撑计划资助项目(2009BAI87B02);国家自然科学基金资助项目(30571875)~~

摘　　要：目的 BMSCs作为雪旺细胞的替代细胞可提高周围神经损伤修复效果,但目前对于神经支架内添加适宜的种子细胞数量未达成共识。研究不同数量BMSCs对大鼠背根神经节(dorsal root ganglion,DRG)生长的影响。方法取4周龄雄性SD大鼠3只,体重80～100 g,体外培养扩增BMSCs。取新生1～2日龄SD大鼠3只,雌雄不限,体重4～6 g,制备DRG。取第3代BMSCs制备BMSCs-生物蛋白胶复合物,按照加入细胞数量的不同,将实验分为A组(1×103个)、B组(1×104个)、C组(1×105个)和D组(0个),并与SD大鼠DRG共培养。48 h后通过形态学观察,神经丝蛋白200和雪旺细胞S-100免疫荧光染色,对SD大鼠DRG轴突生长长度、雪旺细胞迁移距离和轴突面积指数进行定量评价。结果 48 h后可见各组BMSCs生长出多个长突起;雪旺细胞移行和轴突从DRG长出,呈多方向性生长;BMSCs生物蛋白胶具有生物活性且影响DRG生长。A、B、C组DRG轴突生长长度及雪旺细胞迁移距离均明显大于D组(P<0.05),C组小于B组(P<0.05),A、C组间及A、B组间差异无统计学意义(P>0.05)。A、B组轴突面积指数明显大于D组(P<0.05),C组大于D组但差异无统计学意义(P>0.05),A、B、C组间差异无统计学意义(P>0.05)。结论乳鼠DRG体外培养实验,可作为研究周围神经损伤修复的体外模型。不同数量BMSCs生物蛋白胶复合物对DRG生长的影响具有量效关系,为组织工程神经选用合适数量BMSCs提供了理论依据。Objective Bone marrow mesenchymal stem cells（BMSCs）,as replacement cells of Schwann cells,can increase the effect of peripheral nerve repair.However,it has not yet reached any agreement to add the appropriate number of seeded cells in nerve scaffold.To investigate the effect of different number of BMSCs on the growth of rat dorsal root ganglia（DRG）.Methods Three 4-week-old Sprague Dawley（SD） rats（weighing 80-100 g） were selected to isolate BMSCs,which were cultured in vitro.Three 1-to 2-day-old SD rats（weighing 4-6 g） were selected to prepare DRG.BMSCs at passage 3 were used to prepare BMSCs-fibrin glue complex.According to different number of BMSCs at passage 3 in fibrin glue,experiment was divided into group A（1 × 103）,group B（1 × 104）,group C（1 × 105）,and group D（0,blank control）,and BMSCs were co-cultured with rat DRG.The axon length of DRG,Schwann cell migration distance,and axon area index were quantitatively evaluated by morphology,neurofilament 200,and Schwann cells S-100 immunofluorescence staining after cultured for 48 hours.Results Some long cell processes formed in BMSCs at 48 hours;migration of Schwann cells and axons growth from the DRG were observed,growing in every direction.BMSCs in fibrin glue had the biological activity and could effect DRG growth.The axon length of DRG and Schwann cell migration distance in groups A,B,and C were significantly greater than those in group D（P〈 0.05）.The axon length of DRG and Schwann cell migration distance in group C were significantly less than those in group B（P〈 0.05）,but there was no significant difference between group A and group C,and between group A and group B（P〈 0.05）.The axon area index in groups A and B was significantly greater than that in group D（P〉 0.05）,but there was no significant difference between group C and group D（P 〉0.05）;there was no significant difference in groups A,B,and C（P 〉0.05）.Conclusion In vitro study on DRG culture experiments is an ideal objective neur

关 键 词：背根神经节 BMSCS 神经再生 组织工程神经 大鼠 

分 类 号：R745[医药卫生—神经病学与精神病学]