私たちは、ヘルスリサーチ（CIHR）のカナダの研究所からのこの仕事のための資金援助に感謝し、貢献のためKrembilファミリー財団、神経修復におけるハルバート議長と再生、フィリップとペギーDeZwirek、とゴードンヤオは図2を。クラウスZweckbergerは「ドイツForschungsgesellschaft」（DFG）からの助成金によって賄われていた。

注：以下の実験プロトコルは、大学健康ネットワーク（トロント、カナダ）の動物管理委員会によって承認され、動物のケアのカナダ人評議会により調製し、実験動物の世話と使用するためのガイドに設立されたポリシーに従っているた。 1.子宮頸動脈瘤クリップ挫傷/圧縮モデル<ol><li>手術オートクレーブ器の前および70％アルコール浴に器具を置くことによって全体surgcial手順中に無菌状態を維持する。 </li><li> （1：1）酸素との組合せ（O 2）、亜酸化窒素（N 2 O）とのWistarラット（250〜270グラム）を麻酔し、そして1.8から2.2パーセントイソフルランおよびガス麻酔マスクを介して自発呼吸をサポートする。麻酔の誘導のために1分間、5％イソフルランで開始し、その後減少させる。手術を開始する前に、痛みを伴う刺激を与えることにより、麻酔の深さを制御する（ 例えば 。足で）。脂肪軟膏を適用します目に乾燥を避けるために、後続の感染を防止することができる。 </li><li>発熱クッション（37℃）にラットを入れて、定位フレームに頭部を固定する。 </li><li>頸椎周りの手術部位を剃るとポビドンヨードおよび70％アルコールで消毒する。 </li><li>胸椎椎体2（T2）の著名な突起に到達頸椎（C2）から脊柱上記正中切開を行います。 </li><li>頭尾方向（出血を避けるために）直接正中線上の椎骨の筋肉の外側の層を通って切断し、さらにあなたが棘突起とラミナに到達するまで、ぶっきらぼうに深い筋肉層を分析。リトラクターを挿入します。 </li><li>椎弓切除術の対象となるレベルを観察する椎骨本体T2の著名な棘突起を方向付ける。識別と選択したラミナの微小外科調製後、ラミナと棘突起を緩めligamenti flavaeを切断。最後に、C脊髄に対する骨クリッパー横にラミナを通じてUTと脊髄自体のいずれかの圧縮を避け、静かにそれらを削除します。 注：ほとんどの一般的なレベルは、C5 / 6、C6 / 7、またはC7 / T1である。周術期死亡率は損傷レベル吻側より増加します。脊椎傍静脈洞から出血することは一般的であり、スポンジで慎重な圧縮によって対処することができます。 </li><li>コー​​ドを傷つけるためにクリップを挿入する前に、（C5 / 6特にレベルで）クリッピングからそれらを惜しまする新興の神経根を識別する。 </li><li>スムーズなクリップ誘導を確実にするために、フックで椎体の背側から腹側硬膜を緩め、クリップの廊下を準備します。 </li><li>最後に、オープンなクリップを挿入し、それが挫傷を達成するためにシャット（迅速な閉鎖を）スナップましょう。クリップ閉鎖力およびクリップ閉鎖の期間は外傷強度および圧縮の程度を決定する。一般的に使用されるgおよびclippi 15-35間のクリップ力である例えば 1分のNG時間。 （ 図1）。 </li><li>クリップを除去した後、2層の筋肉を適応させ、創傷を閉じる。 </li><li>麻酔を停止し、それが胸骨横臥のために十分な意識を取り戻すまで、動物はあなたの継​​続的な監視下に目を覚ますてみましょう。最後に、一つのケージにラットを入れて、術後の治療ガイドラインに従ってください。 </li><li>動物が傷害のこのタイプの重症度に苦労するので、あなたは、術後の治療に特別な注意を払う必要があります。 <ol><li> （それぞれ、3日間と5日間ブプレノルフィン及びメロキシカムおよび臨床症状に応じて）鎮痛剤を投与する。 </li><li> 3日間の追加の生理食塩水を皮下に与え（1日2回5〜10ミリリットル（ml）で）。 </li><li> 2日前に飲料水で抗生物質を提供し、7日後に手術するまで、（ 例えばモキシフロキサシン） </li><li>膀胱機能の回復が一定になるまで、膀胱を1日2〜3回絞るLY明らか。 </li><li>少なくとも1日に1回神経学的欠損と手術動物の生理学的状態を観察します。 </li></ol></li></ol> 2.注入のSAPとのNPC（損傷後14日間） <ol><li> 1.1で説明したように麻酔を誘導する。 1.3に、定位フレームにラットの頭部を固定し、ステッチまたは創傷クリップを削除し、傷やポビドンヨードおよび70％アルコールで手術領域を消毒する。 </li><li>慎重に、脊椎傍の筋肉を解剖リトラクターを挿入し、硬膜から微視的に瘢痕組織を除去し、病変部位を再公開します。 </li><li>細胞外マトリックスゲルを実行する（w / v）の1％の濃度でSAPを準備する。 QL6のSAPは、生理学的に適合性のpHを持っており、注射の前にバッファリングする必要はありません。脊髄におけるSAPの可視化のため、QL6（QL6-FITC）の蛍光誘導体を使用しています。 </li><li> SAPは（2回に分けて分布する病変の中心に5マイクロリットル（μL）、それぞれを注入正中線の2.5μlの二国間。マイクロガラスキャピラリー（100マイクロメートル（μm）の外径（OD））との定位フレームに接続ハミルトンシリンジを使用してください。鋭い針の先端で慎重に硬膜を開き、心に傷を負った脊髄に定位的に2ミリメートル（mm）をガラスキャピラリーを挿入します。 </li><li>ボリュームの1/3を注入した後、1.5mmの深さに針を削除し、さらに3分の1〜1ミリ後に。全体積の注入後、シリンジを除去する前に、ゲル形成を安定化させるために5分間待つ。 </li><li>のNPCを生成するには、大人のDsRedマウス（または、緑YFP陽性マウス）を使用し、隔離し、paraventricalゾーン4,18,21からそれらを養う。 </li><li>細胞懸濁液中〜90％の生細胞の存在を示す、トリパンブルー染色によるのNPCの生存率を評価。成長培地中で細胞（50×103生細胞/μl）を希釈した後、細胞移植のためにそれらを使用。 </li><li> 4 2μLを作る（8μlの合計VOLUME、吻側と損傷部位の尾2ミリメートルで、両側）髄腔内注射を4×10 5のNPCを含む。硬膜の開口した後、1.5ミリメートル脊髄の背側表面の下にハミルトンマイクロガラス管を挿入し、細胞懸濁液に2μlを注入する。約0.5μL/分（分）で注入速度を選択してください。 </li><li>コー​​ドのうち、各注入の最後には、キャピラリーの除去の前に、組織は新しい細胞容積に対応するために、ストレッチ可能にする少なくとも1分間待ちます。 （ 図2） </li></ol>硬膜下の3。移植は、成長因子の応用のためのポンプ<ol><li>細胞の生存を支持する成長因子と、脳脊髄液（CSF）を豊かにするために、0.5μlの/ hrの希釈率、0.04のカテーテルの直径は、7~14日を通してサブdurally成長因子を希釈微小浸透圧ポンプを使用してセンチOD、そして100μlのリザーバボリューム。 </li><li>好適な成長因子を選択する（例えば、脳由来成長因子（BDGF）、上皮成長因子（EGF）、線維芽細胞増殖因子（FGF））、ポンプ6~10時間、移植前に充填し、37℃の水浴中でポンプを平衡化する。 </li><li>すぐにNPCの注射の後、ポンプを配置するために皮下凹部を準備する。好ましい位置は、ポンプ自体に起因する主要な地元の不快感を回避thoraco-腹部の横方向の両サイドからである。 </li><li>成長因子の最高濃度を得るために、カテーテルの開放先端が病変部位の近くに終了することを確認してください。そのため、隣接する上側または下のレベルのスキップ椎弓切除を行う。 SCIはC7 / T1である場合、C5の小さな椎弓切除を行う。 </li><li> 、皮下凹部にポンプを入れて必要な長さにカテーテルを短縮し、あらゆる動きに関連した転位を避け脊椎傍筋肉上のいくつかの縫合糸（6.0）で固定します。 </li><li>鋭い先端で（C5で、 例えば ）硬膜を開いた後針の、硬膜下腔にカテーテルを導入し、それが抵抗なくしてコードを傷つけることなく尾側方向に滑空してみましょう。 例えば C6のスキップされたラミナは、カテーテルの固定及び安定化の追加のポイントとして機能します。 </li><li>カテーテルがスムーズに実行され、フォールドしないことを確認してください。 </li><li>縫合糸またはクリップで閉じる層によって、筋肉、および皮膚、。 （ 図3） </li><li>麻酔を停止し、それが胸骨横臥のために十分な意識を取り戻すまで、動物はあなたの継​​続的な監視下に目を覚ますてみましょう。最後に、一つのケージにそれらを戻すと術後治療ガイドラインに従ってください。 </li><li>にもかかわらず、動物は、この時点で部分的に回復したかもしれない特別な術後治療を提供する。 <ol><li> （それぞれ、3日間と5日間Buprenorthineとメロキシカムおよび臨床症状に応じて）鎮痛剤を投与する。 </li><li> 2日前まで、水のボトルに抗生物質を提供する7日後に手術（ 例えばモキシフロキサシン）。 </li><li>それでも、必要に応じて、膀胱を圧迫続行。 </li><li>ラットは、脱水表示された場合、追加の皮下流体を与えます。 </li><li>少なくとも1日に1回手術動物の神経学的機能と生理的状態を観察してください。 </li><li> 2日前にNPCの注入を免疫抑制治療を管理し、7日後にtransplanation（ミノサイクリン）と犠牲になるまで（サンディミュン）まで、それぞれ。 </li></ol></li></ol> 4.組織の評価<ol><li>観測時間の終わり（ 例えば SCI後4週間）深麻酔（5％イソフルラン2-3分間）での動物を生け贄に捧げると4冷たい150ミリリットルに続く50ミリリットルの冷（4℃）生理食塩水で経それらを灌流0.1 Mリン酸緩衝生理食塩水（PBS）中の％パラ。 </li><li>脊髄を取り外し、24時間0.1 Mリン酸緩衝生理食塩水（PBS）中4％パラホルムアルデヒド中に入れた。 </li><li&gt; 48時間凍結保護のために25％のショ糖溶液中で脊髄を置く。 </li><li> &lt;厚さ30μmの縦凍結切片を提供します。 </li><li>背景使用DAPI（：1,000 1）を提供するすべての細胞核の免疫組織化学染色のために。 DsRedは正のNPCが赤く表示され、QL-6 FITCは緑表示され、両方のは、このように特別に染色されていない必要がある。 （ 図4）。 </li><li>走査電子顕微鏡（SEM）用のサンプルは、2時間、4℃にてグルタルアルデヒドに浸し、5分間のエタノール10％の増分ステップでゆっくりとそれらを脱水し、1時間加圧された液体CO 2サイフォンに配置しましょう。スパッターコーターを使用して金で被覆足場。日立S-3400N走査型電子顕微鏡で画像を撮影する。 （ 図5） </li></ol>

<ol>
	<li>Sekhon, L. H., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology,+demographics,+and+pathophysiology+of+acute+spinal+cord+injury.">Epidemiology, demographics, and pathophysiology of acute spinal cord injury.</a> Spine (Phila Pa 1976). 26, S2-S12 (2001).</li><li>Fehlings, M. G., Tator, C. H., Linden, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationships+among+the+severity+of+spinal+cord+injury,+motor+and+somatosensory+evoked+potentials+and+spinal+cord+blood+flow).">The relationships among the severity of spinal cord injury, motor and somatosensory evoked potentials and spinal cord blood flow).</a> Electroencephalogr Clin Neurophysiol. 74, 241-259 (1989).</li><li>Thuret, S., Moon, L. D., 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=Therapeutic+interventions+after+spinal+cord+injury.">Therapeutic interventions after spinal cord injury.</a> Nat Rev Neurosci. 7, 628-643 (2006).</li><li>Iwasaki, M., Wilcox, J. T., Nishimura, Y., Zweckberger, K., Suzuki, H., Wang, J., Liu, Y., Karadimas, S. K., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+effects+of+self-assembling+peptide+and+neural+stem/progenitor+cells+to+promote+tissue+repair+and+forelimb+functional+recovery+in+cervical+spinal+cord+injury.">Synergistic effects of self-assembling peptide and neural stem/progenitor cells to promote tissue repair and forelimb functional recovery in cervical spinal cord injury.</a> Biomaterials. 35, 2617-2629 (2014).</li><li>Holmes, T. C., de Lacalle, S., Su, X., Liu, G., Rich, A., Zhang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extensive+neurite+outgrowth+and+active+synapse+formation+on+self-assembling+peptide+scaffolds.">Extensive neurite outgrowth and active synapse formation on self-assembling peptide scaffolds.</a> Proc Natl Acad Sci U S A. 97, 6728-6733 (2000).</li><li>Dong, H., Paramonov, S. E., Aulisa, L., Bakota, E. L., Hartgerink, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembly+of+multidomain+peptides:+balancing+molecular+frustration+controls+conformation+and+nanostructure.">Self-assembly of multidomain peptides: balancing molecular frustration controls conformation and nanostructure.</a> J Am Chem Soc. 129, 12468-12472 (2007).</li><li>Rivlin, A. S., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+duration+of+acute+spinal+cord+compression+in+a+new+acute+cord+injury+model+in+the+rat.">Effect of duration of acute spinal cord compression in a new acute cord injury model in the rat.</a> Surg Neurol. 10, 38-43 (1978).</li><li>Poon, P. C., Gupta, D., Shoichet, M. S., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clip+compression+model+is+useful+for+thoracic+spinal+cord+injuries:+histologic+and+functional+correlates.">Clip compression model is useful for thoracic spinal cord injuries: histologic and functional correlates.</a> Spine (Phila Pa 1976). 32, 2853-2859 (2007).</li><li>Fehlings, M. G., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationships+among+the+severity+of+spinal+cord+injury,+residual+neurological+function,+axon+counts,+and+counts+of+retrogradely+labeled+neurons+after+experimental+spinal+cord+injury.">The relationships among the severity of spinal cord injury, residual neurological function, axon counts, and counts of retrogradely labeled neurons after experimental spinal cord injury.</a> Exp Neurol. 132, 220-228 (1995).</li><li>Joshi, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+a+novel,+graded+model+of+clip+compressive+spinal+cord+injury+in+the+mouse:+Part+2.+Quantitative+neuroanatomical+assessment+and+analysis+of+the+relationships+between+axonal+tracts,+residual+tissue,+and+locomotor+recovery.">Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 2. Quantitative neuroanatomical assessment and analysis of the relationships between axonal tracts, residual tissue, and locomotor recovery.</a> J Neurotrauma. 19, 191-203 (2002).</li><li>Joshi, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+a+novel,+graded+model+of+clip+compressive+spinal+cord+injury+in+the+mouse:+Part+1.+Clip+design,+behavioral+outcomes,+and+histopathology.">Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 1. Clip design, behavioral outcomes, and histopathology.</a> J Neurotrauma. 19, 175-190 (2002).</li><li>Cigognini, D., Satta, A., Colleoni, B., Silva, D., Doneg&#224;, M., Antonini, S., Gelain, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+early+and+late+effects+into+the+acute+spinal+cord+injury+of+an+injectable+functionalized+self-assembling+scaffolds.">Evaluation of early and late effects into the acute spinal cord injury of an injectable functionalized self-assembling scaffolds.</a> PLoS One. 6 (5), e19782 (2011).</li><li>Hou, T., Wu, T., Wang, L., Liu, Y., Li, M., Long, Z., Chen, H., Li, Y., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+prostheses+fabricated+with+motor+neurons+seeded+in+self-assembling+peptides+promotes+partial+functional+recovery+afters+spinal+cord+injury+in+rats.">Cellular prostheses fabricated with motor neurons seeded in self-assembling peptides promotes partial functional recovery afters spinal cord injury in rats.</a> Tissue eng Part A. 18 (9-10), (2012).</li><li>Gelain, F., Cigognini, D., Caprini, A., Silva, D., Colleoni, B., Doneg&#224;, M., Antonini, S., Cohen, B. E., Vescovi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+bioactive+motifs+and+their+use+in+functionalized+self-assembling+peptides+for+NPC+differentiation+and+neural+tissue+engineering.">New bioactive motifs and their use in functionalized self-assembling peptides for NPC differentiation and neural tissue engineering.</a> Nanoscale. 4 (9), 2946-2957 (2012).</li><li>Liu, Y., Ye, H., Satkunendrarajah, K., Yao, G. S., Bayon, Y., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+self-assembling+peptide+reduces+glial+scarring,+attenuates+post-traumatic+inflammation+and+promotes+neurological+recovery+following+spinal+cord+injury.">A self-assembling peptide reduces glial scarring, attenuates post-traumatic inflammation and promotes neurological recovery following spinal cord injury.</a> Acta Biomater. 9, 8075-8088 (2013).</li><li>Rosner, J., Avalos, P., Axosta, F., Liu, J., Drazin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+for+cell+therapy+combined+with+growth+factors+in+spinal+cord+injury.">The potential for cell therapy combined with growth factors in spinal cord injury.</a> Stem Cell Int. , 826754 (2012).</li><li>Lu, P., Wang, Y., Graham, L., McHale, K., Gao, M., Wu, D., Brock, J., Blesch, A., Rosenzweig, E. S., Havton, L. A., Zheng, B., Conner, J. M., Marsala, M., Tuszynsky, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+distance+growth+and+connectivity+of+neural+stem+cells+after+severe+spinal+cord+injury.">Long distance growth and connectivity of neural stem cells after severe spinal cord injury.</a> Cell. 150, 1265-1273 (2012).</li><li>Karimi-Abdolrezaee, S., Schut, D., Wang, J., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chondrioitinase+and+grwoth+factors+enhance+activation+and+oligodendrocyte+differentiation+of+endogenous+neural+precursor+cells+after+spinal+cord+injury.">Chondrioitinase and grwoth factors enhance activation and oligodendrocyte differentiation of endogenous neural precursor cells after spinal cord injury.</a> PLoS One. 7 (5), e37589 (2012).</li><li>Awad, B. I., Carmody, M. A., Steinmetz, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+role+of+growth+factors+in+the+management+of+spinal+cord+injury.">Potential role of growth factors in the management of spinal cord injury.</a> World Neurosurg. (13), 1875-8750 (2013).</li><li>Kojima, A., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrathecal+administration+of+epidermal+growth+factor+and+fibroblast+growth+factor+2+promotes+ependymal+proliferation+and+functional+recovery+after+spinal+cord+injury+in+adult+rats.">Intrathecal administration of epidermal growth factor and fibroblast growth factor 2 promotes ependymal proliferation and functional recovery after spinal cord injury in adult rats.</a> J Neurotrauma. 19 (2), 223-238 (2002).</li><li>Karimi-Abdolrezaee, S., Eftekharpour, E., Wang, J., Cindi, M. M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+trasplantation+of+adult+neural+presursor+cells+promotes+remyelination+and+functional+neurological+recovery+after+spinal+cord+injury.">Delayed trasplantation of adult neural presursor cells promotes remyelination and functional neurological recovery after spinal cord injury.</a> J Neurosci. 26 (13), 3377-3389 (2006).</li><li>Burdick, J. A., Ward, M., Liang, E., Young, M. J., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+neurite+outgrowth+by+neurotrophins+delivered+from+degradable+hydrogels.">Stimulation of neurite outgrowth by neurotrophins delivered from degradable hydrogels.</a> Biomaterials. 27, 452-459 (2006).</li></ol>

Covidien supplied the QL6 SAP for this study.

このプロトコルは、ラットにおいて、子宮頸部傷害モデルを実行するために読者を可能にし、子宮頸SCI後のより良い回復を促進するのSAPとのNPCとの併用治療アプローチを使用するために開発された。 挫傷と圧縮 - 、したがって、模倣最高の臨床症状 - 特にそのような（半）-transectionモデルまたは重量の低下と脳震盪モデルなどの他の頸椎外傷モデル、と比較して、クリ?...

脊髄損傷の50％以上が頸椎に関連している。脊髄の初期挫傷を、その後、継続的な圧縮は、骨折、出血または組織の腫脹によって引き起こされる臨床設定つの主要な病態生理学的メカニズムに記載されている。 動脈瘤クリップ挫傷/圧縮モデルを模倣両方の病態生理学的メカニズム：クリップをスナップする挫傷を生成し、クリッピングの持続時間は、骨折、出血または組織によって引き起こされる臨床現場での圧縮は、最後の有意な長い腫脹こと失点、圧縮成分を表す。使用される動脈瘤クリップを正確かつ再現性のクリップ力を保証するリングばねによって変更される。特に、ヘミ離断または挫傷モデル、この動脈瘤クリップモデルを模倣最良の臨床設定と比較して。頸椎INJと胸部外傷の患者は麻痺に苦しむ一方で、ほとんどの患者uriesは四肢麻痺と完全に依存しています。頸髄の解剖学的構造は、しかしながら、胸椎または腰椎と比較して有意差を示し、従って、このプロトコルに特に対処される。 髄内空洞および組織瘢痕化の開発は回復と再生のための障害となっている。これらの障害の足場材料の使用を克服するための有望なアプローチである。自己組織化ペプチドは、病変の震源地に直接注入することができます。そこで彼らは、空洞を埋めるナノファイバー足場に集合し、炎症および組織瘢痕を減らすことによって抑制環境を改善する。剛性材料は、注入時の脊髄のかなりの損傷を引き起こす一方で、流体ペプチドが安全かつ深刻なダメージを与えることなく、追加注入することができる。 したがって、幹細胞移植前に自己集合性ペプチドとの阻害性環境の改善、細胞のiをサポートntegration頸部脊髄損傷後の分化し、最終的に、機能回復、。

<table border="0" cellpadding="0" cellspacing="0" width="541">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:273px;">Name</td>
			<td style="width:268px;">Company</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Aneurysmal clip</td>
			<td>SharpTech</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical microscope</td>
			<td>Leica</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro injection system</td>
			<td>World Precision Instruments, Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small animal stereotaxic instrument</td>
			<td>David Kopf Instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hamilton syringe</td>
			<td>Hamilton company</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Subdural pumps</td>
			<td>Alzet osmotic micro pump 1007D</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical instrument</td>
			<td>Fine Science tools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane USP</td>
			<td>Pharmaceutical Partners of Canada Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.9% Sodium Chloride injection USP</td>
			<td>Baxter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7.5% Povidone iodine</td>
			<td>Purdue Pharma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% Isopropyl alcohol USP</td>
			<td>GreenField Ethanol Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">QL6 SAP</td>
			<td>Covidien</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.4% Trypan blue</td>
			<td>Gibco</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platelet-Derived Growth Factor (PDGF)</td>
			<td>Sigma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Epidermal Growth Factor (EGF)</td>
			<td>Sigma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fibroblast Growth Factor (FGF)</td>
			<td>Sigma</td>
		</tr>
	</tbody>
</table>

synergetic use of neural precursor cells and self-assembling peptides in experimental cervical spinal cord injury

脊髄損傷（SCI）は、患者とその家族のために深刻な神経学的障害や、心理的、経済的、社会的影響を引き起こす。臨床的には、SCIの50％以上が頸椎1に影響を与える。主な傷害の結果として、炎症、アポトーシス、および脱髄を含む二次メカニズムのカスケードは、最終的に髄内空洞2,3の組織瘢痕化と発展につながる発生する。どちらも、細胞移植、統合、および再生の物理的および化学的障壁を表す。そのため、抑制性の環境を成形し、細胞移植と再生のための支援環境を作成するための空洞を埋めることは有望な治療ターゲット4である。ここで、動脈瘤クリップを用いて、子宮頸部SCIの挫傷/圧縮モデルが記載されている。完全な離断または脊髄の破裂は稀であるため、このモデルは、より臨床的に関連する他の実験モデルよりも長い。またcomparisoで重量ドロップモデルにN、特にダメージで背の列、脊髄の周圧縮が有利表示されます。クリップ閉鎖力および持続時間は、異なる損傷の重症度を達成するように調整することができる。リングばねは、クリップ力の正確なキャリブレーションと恒常性を容易にします。生理的条件下で、合成自己集合性ペプチド（SAP）ナノファイバーに自己集合し、したがって、SCI 5における適用のために魅力的。それらは脊髄に対する損傷を最小限に病変に直接注射することができる。 SAPは、髄内の空洞を埋めるため、再生治療のために、損傷した脊髄を装備する足場を建てる生体適合性構造体である。急性期後6 0.14日SCI後中性pHでK2（QL）6K2（QL6） ら Lドンによって導入された新規のSAPである。6他のペプチドと比較して、QL6自己集合βシートに、SAPの病変および神経前駆細胞（NPC）の中心に注入される麟蹄である隣接する背側の列にCTED。細胞の生存をサポートするために、移植は、7日間の浸透マイクロポンプによって成長因子の連続的な硬膜下投与と組み合わされる。

<html>上記の手順を実行するときは、。SAP足場空洞を架橋および阻害環境の改善、より少ない組織瘢痕及びNPCの生存の増加を提供し得る。図4になることで得られたラットの脊髄の縦断面を示している損傷部位SCI後6週間とQL6 SAPの注入とNPC移植後4週間。 QL6ペプチドは成功裏に震源地で集約され、コード内に注入し、周辺部で吻側 - 尾側に拡散された。電子顕微鏡図5に、...

Watch this Scientific Journal Video about Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury at JoVE.com

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

実験的子宮頸脊髄損傷における神経前駆細胞と自己組織化ペプチドの相乗使用

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

synergetic-use-neural-precursor-cells-self-assembling-peptides

Research

JoVE Journal

Medicine

9.1K ビュー.  University Health Network, Krembil Neuroscience Centre. Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established. 

ビデオ: Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

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

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

筋萎縮性側索硬化症や外傷性脊髄損傷では頚椎前角をターゲットとするための髄腔内細胞移植

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

医学

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

ラット脊髄損傷後の急性および慢性の触覚官能検査

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

無限の地平線インパクターを用いた片側頚髄損傷の打撲のモデル

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

慢性脊髄損傷患者で呼吸器モータ制御評価（RMCA）を使用して呼吸筋活動の評価

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

ポスト病変呼吸神経可塑性を研究するための子宮頸脊髄損傷のマウスモデル

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

外傷性脳損傷のための制御皮質衝撃モデル

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

ラジオ遠隔測定システムは、脊髄離断および胚性神経細胞移植片幹ラットの心臓血管機能を監視する

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

脊髄損傷の実験的挫傷モデルにおける神経幹細胞移植

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

脊髄圧縮傷害の校正済み鉗子モデル

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...