我想感谢：莱波雷的所有成员，Maragakis和罗斯坦有益的讨论实验室;美国和克雷格H.尼尔森基金会拨款瘫痪退伍军人。

<ol>
	<li>Lane, M. A., Fuller, D. D., White, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+recovery+following+high+cervical+hemisection.">Respiratory recovery following high cervical hemisection.</a> Trends in neurosciences. 31, 538-538 (2008).</li><li>Kaplan, L. M., Hollander, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+dysfunction+in+amyotrophic+lateral+sclerosis.">Respiratory dysfunction in amyotrophic lateral sclerosis.</a> Clin. Chest. Med. 15, 675-675 (1994).</li><li>Miller, R. G., Rosenberg, J. A., Gelinas, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Practice+parameter:+the+care+of+the+patient+with+amyotrophic+lateral+sclerosis+(an+evidence-based+review):+report+of+the+Quality+Standards+Subcommittee+of+the+American+Academy+of+Neurology:+ALS+Practice+Parameters+Task+Force.">Practice parameter: the care of the patient with amyotrophic lateral sclerosis (an evidence-based review): report of the Quality Standards Subcommittee of the American Academy of Neurology: ALS Practice Parameters Task Force.</a> Neurology. 52, 1311-1323 (1999).</li><li>Mitsumoto, H., Chad, D. A., Pioro, E. P., Davis, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Amyotrophic lateral sclerosis. , (1998).</li><li>Tandan, R., Bradley, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amyotrophic+leteral+sclerosis:+Part+2.+Etiopathogenesis.">Amyotrophic leteral sclerosis: Part 2. Etiopathogenesis.</a> Annals of neurology. 18, 419-419 (1985).</li><li>Bruijn, L. I., Miller, T. M., Cleveland, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unraveling+the+mechanisms+involved+in+motor+neuron+degeneration+in+ALS.">Unraveling the mechanisms involved in motor neuron degeneration in ALS.</a> Annu Rev Neurosci. 27, 723-723 (2004).</li><li>Rosen, D. R., Siddique, T., Patterson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+in+Cu/Zn+superoxide+dismutase+gene+are+associated+with+familial+amyotrophic+lateral+sclerosis.">Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis.</a> Nature. 362, 59-59 (1993).</li><li>Bruijn, L. I., Becher, M. W., Lee, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ALS-linked+SOD1+mutant+G85R+mediates+damage+to+astrocytes+and+promotes+rapidly+progressive+disease+with+SOD1-containing+inclusions.">ALS-linked SOD1 mutant G85R mediates damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions.</a> Neuron. 18, 327-327 (1997).</li><li>Gurney, M. E., Pu, H., Chiu, A. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+neuron+degeneration+in+mice+that+express+a+human+Cu,Zn+superoxide+dismutase+mutation.">Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation.</a> Science. 264, 1772-1775 (1994).</li><li>Howland, D. S., Liu, J., She, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+loss+of+the+glutamate+transporter+EAAT2+in+a+transgenic+rat+model+of+SOD1+mutant-mediated+amyotrophic+lateral+sclerosis+(ALS).">Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS).</a> Proc Natl Acad Sci. 99, 1604-1604 (2002).</li><li>Nagai, M., Aoki, M., Miyoshi, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rats+expressing+human+cytosolic+copper-zinc+superoxide+dismutase+transgenes+with+amyotrophic+lateral+sclerosis:+associated+mutations+develop+motor+neuron+disease.">Rats expressing human cytosolic copper-zinc superoxide dismutase transgenes with amyotrophic lateral sclerosis: associated mutations develop motor neuron disease.</a> J. Neurosci. 21, 9246-9246 (2001).</li><li>McDonald, W., Becker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury:+promising+interventions+and+realistic+goals.">Spinal cord injury: promising interventions and realistic goals.</a> Am. J. Phys. Med. Rehabi. I82, S38-S38 (2003).</li><li>Sandrow-Feinberg, H. R., Zhukareva, V., Santi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PEGylated+interferon-beta+modulates+the+acute+inflammatory+response+and+recovery+when+combined+with+forced+exercise+following+cervical+spinal+contusion+injury.">PEGylated interferon-beta modulates the acute inflammatory response and recovery when combined with forced exercise following cervical spinal contusion injury.</a> Experimental neurology. 223, 439-451 (2010).</li><li>Gage, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mammalian+neural+stem+cells.">Mammalian neural stem cells.</a> Science. 287, 1433-1433 (2000).</li><li>Lane, M. A., Lee, K. Z., Fuller, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+circuitry+and+respiratory+recovery+following+spinal+cord+injury.">Spinal circuitry and respiratory recovery following spinal cord injury.</a> Respiratory physiology &#38; neurobiology. 169, 123-123 (2009).</li><li>Lepore, A. C., Rauck, B., Dejea, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+transplantation-based+astrocyte+replacement+is+neuroprotective+in+a+model+of+motor+neuron+disease.">Focal transplantation-based astrocyte replacement is neuroprotective in a model of motor neuron disease.</a> Nature. 11, 1294-1294 (2008).</li><li>Rao, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multipotent+and+Restricted+Precursors+in+the+Central+Nervous+System.">Multipotent and Restricted Precursors in the Central Nervous System.</a> Anat Rec. 257, 137-137 (1999).</li><li>Rao, M. S., Mayer-Proschel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial-+restricted+precursors+are+derived+from+multipotent+neuroepithelial+stem+cells.">Glial- restricted precursors are derived from multipotent neuroepithelial stem cells.</a> Dev Biol. 188, 48-48 (1997).</li><li>Suzuki, M., Tork, C., Shelley, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+disease+onset+and+progression+of+a+rat+model+of+ALS.+Sexual+dimorphism+in+disease+onset+and+progression+of+a+rat+model+of+ALS.">Sexual dimorphism in disease onset and progression of a rat model of ALS. Sexual dimorphism in disease onset and progression of a rat model of ALS.</a> Amyotroph Lateral Scler. 8, 20-20 (2007).</li><li>Lepore, A. C., Haenggeli, C., Gasmi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraparenchymal+spinal+cord+delivery+of+adeno-associated+virus+IGF-1+is+protective+in+the+SOD1G93A+model+of+ALS.">Intraparenchymal spinal cord delivery of adeno-associated virus IGF-1 is protective in the SOD1G93A model of ALS.</a> Brain research. 1185, 256-256 (2007).</li><li>Veldink, J. H., Bar, P. R., Joosten, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+differences+in+onset+of+disease+and+response+to+exercise+in+a+transgenic+model+of+ALS.">Sexual differences in onset of disease and response to exercise in a transgenic model of ALS.</a> Neuromuscul Disord. 13, 737-737 (2003).</li><li>Shumsky, J. S., Lepore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+Neuronal+and+Glial+Restricted+Precursors+into+Contused+Spinal+Cord+Improves+Bladder+and+Motor+Functions,+Decreases+Thermal+Hypersensitivity,+and+Modifies+Intraspinal+Circuitry.">Transplantation of Neuronal and Glial Restricted Precursors into Contused Spinal Cord Improves Bladder and Motor Functions, Decreases Thermal Hypersensitivity, and Modifies Intraspinal Circuitry.</a> J. Neurosci. 25, 9624-9624 (2005).</li></ol>

没有利益冲突的声明。

研究涉及的SOD1 G93A小鼠和大鼠，年龄和性别匹配的一组内的动物，和分发不同群体的同一窝内的动物。最好是使用ALS和SCI模型相同性别的所有动物，因为疾病的进程可能男性和女性之间有所不同，但是，它也可能是有用的男女双方有足够的动物，以检测出可能特定性别的影响，这一现象已报道G93A SOD1的老鼠13和G93A SOD1的老鼠14,15。考虑细胞移植到动物的年?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>的HBSS </td><td> Gibco公司</td><td> 14170 </td></tr><tr><td> 0.05％胰蛋白酶</td><td> Gibco公司</td><td> 25300 </td></tr><tr><td>大豆胰蛋白酶抑制剂（可选） </td><td>西格玛</td><td> T - 6522 </td></tr><tr><td> Acepromazine马来酸（0.7毫克/公斤） </td><td>发酵动物健康</td><td></td></tr><tr><td>氯胺酮（95毫克/公斤） </td><td>道奇堡动物健康</td><td></td></tr><tr><td>甲苯噻嗪（10毫克/公斤） </td><td>拜耳</td><td></td></tr><tr><td> ＃11羽手术刀片</td><td>电子显微镜科学</td><td> 72044-11 </td></tr><tr><td>棉签涂药（6英寸） </td><td>费舍尔</td><td> 23-400-101 </td></tr><tr><td>鼠有齿镊</td><td>精细科学的工具</td><td>鼠：11023-15; 鼠标：11042-08 </td></tr><tr><td>中型弹簧剪刀</td><td>精细科学的工具</td><td> 15012-12 </td></tr><tr><td>微型弹簧剪刀</td><td>精细科学的工具</td><td> 15000-10 </td></tr><tr><td>咬骨钳</td><td>精细科学的工具</td><td>鼠：16121-14; 鼠标：16221-14 </td></tr><tr><td> Microknife </td><td>精细科学的工具</td><td> 10056-12 </td></tr><tr><td>持针</td><td>精细科学的工具</td><td> 12502-14 </td></tr><tr><td>缝合：4-0 </td><td>薇乔</td><td> S - 183 </td></tr><tr><td>主食：9毫米</td><td> Autoclip </td><td> 427631 </td></tr><tr><td>订书机：9毫米（＃203-1000反射） </td><td>世界精密仪器</td><td> 5000344 </td></tr><tr><td>暖水泵（T /泵） </td><td> Gaymar </td><td> P / ñ 07999-000 </td></tr><tr><td>环孢素A：250.0 mg/5.0毫升安瓿</td><td>诺华/ Sandimmune </td><td> NDC 0078-0109-01 </td></tr><tr><td> FK - 506 </td><td>立法会实验室</td><td> F - 4900 </td></tr><tr><td>雷帕霉素</td><td>立法会实验室</td><td> R - 5000 </td></tr><tr><td>注射器</td><td>世界精密仪器</td><td> UMP2 </td></tr><tr><td>微型4 Microsyringe泵控制器</td><td>世界精密仪器</td><td> UMC4 </td></tr><tr><td>微操作机器人</td><td>世界精密仪器</td><td>风筝 - R </td></tr><tr><td> 10.0微升汉密尔顿注射器</td><td>汉密尔顿</td><td> 80030 </td></tr><tr><td>汉密尔顿针：33号，45 °斜角，1英寸</td><td>汉密尔顿</td><td> 7803-05 </td></tr><tr><td>玻璃20.0μLmicrocapillary移液器（可选） </td><td>金布尔</td><td> 71900-20 </td></tr></tbody></table>

intraspinal cell transplantation for targeting cervical ventral horn in amyotrophic lateral sclerosis and traumatic spinal cord injury

呼吸妥协由于膈运动神经元的损失是一个衰弱的后果了大量人力外伤性脊髓损伤（SCI）的例1的比例，是与运动神经元症，肌萎缩硬化症（ALS）2支管患者死亡的最终原因。 ALS是一种破坏性的神经系统疾病的特点是较快的上下运动神经元的变性。屈服于平均2-5年的疾病的患者最终因呼吸麻痹由于膈运动神经元膜片3 innnervation损失的诊断。绝大多数情况下是零星的，而10％的家族形式。大约二十％的家族性病例是在21号染色体 4铜/锌超氧化物歧化酶1（SOD1）基因点突变。转基因小鼠4,5和6大鼠携带突变人类的SOD1基因（G93A，G37R，G86R，G85R）已经产生，并且，尽管存在着其他的动物模型运动神经元的损失，目前该疾病的最高度使用的模型。 脊髓损伤（SCI）是一个异构的条件，造成身体创伤，脊髓功能的结果，根据不同的类型，位置和严重程度的伤害 7 。然而，大约一半人类脊髓损伤病例的影响颈椎的地区，导致呼吸功能障碍衰弱由于膈运动神经元的损失和伤害的降bulbospinal呼吸轴突1。已开发的脊髓损伤动物模型，最常用的和临床相关挫伤8。 各种类的神经前体细胞（NPC的）移植是治疗外伤性中枢神经系统损伤和神经退行性疾病，包括ALS和SCI的一种很有前途的治疗策略，因为更换丢失或功能失调性中枢神经系统的细胞类型，提供神经保护作用的能力，并提供基因因素9利益。 ALS和SCI的动物模型，可以模拟许多这些疾病的临床相关的各个方面，包括膈运动神经元的损失和由此产生的呼吸妥协10,11。为了评估全国人民代表大会为基础的战略，在ALS和SCI这些动物模型的呼吸功能的疗效，必须专门针对蜂窝干预含有治疗膈运动神经元等有关目标的地区。我们提供一个详细的协议为多节段椎管内的NPC移植到颈椎脊髓腹侧灰质神经退行性型号如SOD1的G93A小鼠和大鼠，以及脊髓损伤大鼠和小鼠11。

Watch this Scientific Journal Video about Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury at JoVE.com

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

神经前体移植是一种很有前途的战略，保护和/或更换丢失/功能失调颈椎膈运动神经元在脊髓损伤（SCI）和运动神经元症，肌萎缩边音硬化症（ALS）。我们提供了一个细胞传递到脊髓型颈椎病ALS和SCI的啮齿类动物模型腹角的协议。

椎管内针对肌萎缩性脊髓侧索硬化症和外伤性脊髓损伤颈腹角细胞移植

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

intraspinal-cell-transplantation-for-targeting-cervical-ventral-horn

Research

JoVE Journal

Medicine

20.5K 次观看.  Thomas Jefferson University Medical College. 神经前体移植是一种很有前途的战略，保护和/或更换丢失/功能失调颈椎膈运动神经元在脊髓损伤（SCI）和运动神经元症，肌萎缩边音硬化症（ALS）。我们提供了一个细胞传递到脊髓型颈椎病ALS和SCI的啮齿类动物模型腹角的协议。