脑外科刘教授11月14日课件2(改1).ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,西京医院神经外科,刘卫平 费舟等,干细胞移植治疗脑损伤的,基础及临床研究,The cilinical and basic research of traumatic brain injury by stem cells transplantation,WHAT CAN WE DO NOW,?,大骨瓣减压术,脱水剂,激素,冬眠疗法,高压氧,自由基清除剂,钙离子拮抗剂,神经营养药物,约,10%,轻度损伤患者和,66-100%,中、重度损伤患者遗留有永久的神经功能障碍,疗 效？,中枢神经系统不可再生？,神经干细胞Neural stem cells,，,NSCs,）,自我更新,多向分化,可塑性,迁移性,bFGF,、,EGF,BDNF,、,NT-3,全新的治疗思路！,干细胞治疗中枢神经系统损伤,替代治疗：,移植外源,性干细胞,补充治疗：,激活内源,性干细胞,一,、,外源性性干细胞的 实验研究,胚胎干细胞,(ESC),神经干细胞,(NSC),神经元,(Neuron),ESC,分离培养,ESC,诱导分化,ESC,的实验研究,NSC,的实验研究,Nestin,GABA,NSC,诱导分化,NSC,分离培养,葡萄糖,对神经干细胞,增殖分化,的影响,离子浓度变化,对神经干细胞,凋亡,的影响,干细胞微环境变化,存在问题,更好的干细胞？,1,干细胞来源,2,致瘤性,3,伦理问题,二、,内源性性干细胞的 实验研究,内源性神经干细胞具有以下优势,来源可靠稳定,无伦理问题,无免疫排斥反应,无致瘤性,一,二,三,四,碱性成纤维细胞生长因子,(basic Fibroblast Growth Factor,bFGF),促进血管形成,促进修复再生,促进神经元的功能恢复,生物活性较强的促分裂原,建立创伤性脑外伤模型,采用改良的,Feeney,氏自由落体撞击来制作创伤性脑外伤模型,40g,重的铁质圆柱体自,25cm,高处自由落体撞击,左侧顶骨咬除,5.0mm,5.0mm,显露硬脑膜,脑室内注射,4A,：对照组,(DAB,染色，,100),4B,：对照组,(DAB,染色，,400),4C,：治疗组,(DAB,染色，,100),4D,：治疗组,(DAB,染色，,400),损伤侧海马组织中,BrdU,阳性细胞,BrdU-positive cells at the DG of the hippocampus on the damage side,结果,重型颅脑损伤？,侧脑室室管膜下区,(SVZ),海马齿状回颗粒细胞下层（,DG/SGZ,）,取材方便,纯度高,间充质干细胞,无损害,无伦理学限制,无免疫排斥,生物特性好,脐带间充质,干细胞,(UMSC),骨髓间充质,干细胞,(BMSC),Endogenous FactorsExogenous Factors,体外研究,BMSC,CD105,Nestin,NSE,NSC,neurons,+,动物实验,BrdU(Ipsilateral cerebral cortex,Week 1,100,),BrdU(Ipsilateral cerebral cortex,，,Week 2,100,),BrdU(Ipsilateral cerebral cortex,Week 3,100,),BrdU(Ipsilateral cerebral cortex,，,Week 4,100,),BrdU(Ipsilateral,hippocampus,Week 2,100,),BrdU(Ipsilateral,hippocampus,Week 4,400,),BrdU(Ipsilateral,hippocampus,Week 3,400,),BrdU(Ipsilateral,hippocampus,Week 2,400,),BrdU,BrdU,GFAP,NeuN,Merge,Merge,GFAP(Ipsilateral cerebral cortex),NeuN(Ipsilateral,hippocampus,),Ipsilateral cerebral cortex,Ipsilateral,hippocampus,Neun/,-actin,Neun/,-actin,Horizontal motion,Vertical motion,Tiltboard time,Beam-walking,Behavior Scales,A,：,BMSCs+NGF,B,：,BMSCs C,：,Injury control D,：,Blank Control,BMSCs+NGF,group,BMSCS,group,Injury,control,Blank,control,Time factor,Inter-class factor,F,F,Horizontal motion,39.51,0.47,18.45,0.47,8.55,0.49,103.63,0.47,1798.67,8039.39,Vertical motion,9.89,0.18,5.25,0.18,2.21,0.19,21.80,0.18,263.25,1938.69,Tiltboard time,9.29,0.15,6.02,0.15,2.44,0.16,25.37,0.15,356.03,4328.71,Beam-walking,19.81,0.14,24.64,0.13,25.07,0.14,3.67,0.14,435.33,5232.33,Comparison between groups(,P,),A vs.B,A vs.C,A vs.D,B vs.C,B vs.D,C vs.D,Horizontal motion,0.001,0.001,0.001,0.001,0.001,0.001,Vertical motion,0.001,0.001,0.001,0.001,0.001,0.001,Tiltboard time,0.001,0.001,0.001,0.001,0.001,0.001,Beam-walking,0.001,0.001,0.001,0.001,0.001,0.001,三、,临床实验研究,Progress,Ajou University,South Korea,2005,University of Chile,Chile,2007,Pro-Cardiaco Hospital,Brazil,2006,Montreal,CA-Israel,2008,Boston,USA-Greece,2009,Coma duration,23m,Cause,traffic accident,Diagnosis,vegetable,临床病例,HBMSCs(P1),HBMSCs(P3),CD34,CD105,NeuN,GFAP,Day 2,Day 8,BMSC,临床实验,自体,BMSC,体外扩增,BMSC,移植,BMSC,移植,美国多学科,PVS,研究组,提出的“植物状态”,(1994),评分参考,脑卒中患者临床神经功能,缺损程度评分标准,(1995),神经功能缺失的临床评估,影像学检查,效果评估,神经电生理学检查,功能学评价,移植前（,2008/07/01,）,移植后（,2008/09/08,）,移植后（,2009/03/03,）,干细胞与周围组织建立了突触联系,干细胞分泌各种营养因子,可能的机理？,存在问题,老,年人的骨髓脂肪化,-,来源,少,成,年人骨髓干细胞,-,活性,差,急,性颅脑损伤病人,-,周期,长,数量丰富，易于采集,基因稳定，安全可靠,免疫性低，免疫调节,发育更早，环境单纯,修复损伤，重建功能,UMSC,的特点,强强联合,温总理强调：“北科从事的干细胞研究是生物科技的最前沿,是最有希望超越西方发达国家的领域。正如你们所期盼的，,让一个国家、一个民族在一个大的科学领域领先世界！”,UMSC,移植方案,选择病例（签署知情同意书）,完善相关检查,脐带间充质干细胞分离提纯培养制备鉴定,每周,1,次，共,4,次脐带间充质干细胞治疗。,第,1,次：静脉,210,7,（,30mL,）,;,第,2,次：鞘内,210,7,（,1mL,）,;,第,3,次：鞘内,210,7,（,1mL,）,;,第,4,次，鞘内,210,7,（,1mL,）,治疗后观察、检查、并进行定期随访,定期汇总分析，进行有效性、安全性评估,研究项目获军队专项课题资助,干细胞项目是国家和军队的十二五重大研究课题,谢谢！,