マウス局所脳虚血モデルへの神経堤由来細胞移植の効果の検討

2011 Fiscal Year Annual Research Report

• Project/Area Number: 21591119
• Research Institution: Keio University
• Principal Investigator: 冨田 裕 慶應義塾大学, 医学部, 講師(非常勤) (60276251)
• Keywords: 神経堤細胞 / 脳梗塞 / Confocal顕微鏡 / Matlab software

Research Abstract

当実験系において、脳梗塞モデルマウスに、green fluorescent protein(GFP)を発現する遺伝子改変マウスから得られた新鮮幹細胞などを移植した後、レーザー光(488nm)スキャン共焦点顕微鏡を用いて、単一細胞レベルで細胞を生きたまま長期間に亘り反復画像化可能であることが確かめられた。移植した幹細胞が虚血および虚血周辺領域へと移動(migration)、分裂(differentiation)、分化(prohferation)することが明瞭に示された。
一方、神経堤細胞をGFPで標識するPO-Cre/Floxed-EGFPマウスの骨髄より、フローサイトメトリーを用いてGFP陽性の神経堤由来細胞を回収し、得られた神経堤由来細胞を同様に脳梗塞モデルマウスに脳表より1mmの深さに移植した場合には、共焦点顕微鏡により頭窓を通して移植細胞のその後の動態を観察しても、必ずしも個々の細胞レベルまでは移動、分裂、分化の動態が明瞭には観察されないケースもあった。採取され得る移植細胞の数が極端に少ない点が今後の課題として浮かび上がった。また、共焦点顕微鏡では、脳皮質より約100μmの深さまでの観察が可能であるが、より脳深部における細胞の挙動を観察する実験系を今後確立し、さらに詳細に比較検討していく必要性が示唆された。
血栓溶解療法を除くと脳梗塞に対する完全に有効な治療法はいまだ存在せず、さらなる基礎研究の進歩が期待され、近年、再生医療として脳虚血後の骨髄幹細胞移植による治療効果が注目されている。移植する幹細胞の種類(分化・未分化)、数、移植の時期、投与方法などさまざまな課題が残されているが、今回マウス脳梗塞モデルにて、細胞移植後、生きたまま個々の細胞レベルまで長期間反復観察できる実験系を完全に確立したことは、今後の研究発展に大きく寄与していくことと考えている。

Research Products

• [Journal Article] Suppressive effect of chronic peroral topiramate on potassium-induced cortical spreading depression in rats (2012)
  • Author(s): Unekawa M, Tomita Y, Toriumi H, Suzuki N
  • Journal Title: Cephalalgia Volume: (in press)
  • Peer Reviewed
• [Journal Article] Sustained decrease and remarkable increase in red blood cell velocity in intraparenchymal capillaries associated with potassium-induced cortical spreading depression (2012)
  • Author(s): Unekawa M, Tomita M, Tomita Y, Toriumi H, Suzuki N
  • Journal Title: Microcirculation Volume: 19(2) Pages: 166-174
  • Peer Reviewed
• [Journal Article] Repeated Longitudinal in vivo Imaging of Neuro-glio-vascular Unit at the Peripheral Boundary of Ischemia in Mouse Cerebral Cortex (2012)
  • Author(s): Masamoto K, Tomita Y, Toriumi H, Unekawa M, Takuwa H, Itoh Y, Suzuki N, Kanno I
  • Journal Title: Neuroscience Volume: (in press)
  • Peer Reviewed
• [Journal Article] Long-term, repeated measurements of mouse cortical microflow at the same region of interest with high spatial resolution (2011)
  • Author(s): Tomita Y, Pinard E, Tran-Dinh A, Schiszler I, Kubis N, Tomita M, Suzuki N, Seylaz J
  • Journal Title: Brain Research Volume: 1372 Pages: 59-69
  • Peer Reviewed
• [Journal Article] Developmental and circulatory profile of the diploic veins (2011)
  • Author(s): Toriumi H, Shimizu T, Shibata M, Unekawa M, Tomita Y, Tomita M, Suzuki N
  • Journal Title: Microvascular Research Volume: 81 Pages: 97-102
  • Peer Reviewed
• [Journal Article] Oscillating neuro-capillary coupling during cortical spreading depression as observed by tracking of FITC-labeled RBCs in single capillaries (2011)
  • Author(s): Tomita M, Tomita Y, Unekawa M, Toriumi H, Suzuki N
  • Journal Title: NeuroImage Volume: 56(3) Pages: 1001-1010
  • Peer Reviewed
• [Journal Article] Capillary remodeling and collateral growth without angiogenesis after unilateral common carotid artery occlusion in mice (2011)
  • Author(s): Guo H, Itoh Y, Toriumi H, Yamada S, Tomita Y, Hoshino H, Suzuki N
  • Journal Title: Microcirculation Volume: 18(3) Pages: 221-227
  • Peer Reviewed
• [Presentation] Changes in diameter of cerebral microvessels and blood flow associated with potassium-induced cortical spreading depression in anesthetized mice (2012)
  • Author(s): 畝川美悠紀, 冨田裕, 鳥海春樹, 海老根妙子, 正本和人, 伊藤義彰, 菅野巌, 鈴木則宏
  • Organizer: 第37回日本微小循環学会総会
  • Place of Presentation: 盛岡
  • Year and Date: 2012-03-16
• [Presentation] K^+による大脳皮質性拡延性抑制誘発時の局所脳血流と軟膜動脈血管口径の変化 (2011)
  • Author(s): 畝川美悠紀, 冨田裕, 鳥海春樹, 正本和人, 伊藤義彰, 菅野巌, 鈴木則宏
  • Organizer: 第23回日本脳循環代謝学会総会
  • Place of Presentation: 東京
  • Year and Date: 2011-11-04
• [Presentation] マウス一側総頸動脈閉塞後に観察された血管新生を伴わない毛細血管の動脈化と側副血行路の発達:Capillary arterialization and collateral growth without angiogenesis after carotid artery occlusion in mice (2011)
  • Author(s): Guo H, Itoh Y, Toriumi H, Yamada S, Tomita Y, Hoshino H, Suzuki N
  • Organizer: 第36回日本脳卒中学会総会
  • Place of Presentation: 京都
  • Year and Date: 2011-07-30
• [Presentation] Effect of injection of artificial rbcs on murine hemorrhagic hypotension model (2011)
  • Author(s): Tomita Y, Unekawa M, Toriumi H, Masamoto K, Sakai H, Tsuchida E, Horinouchi H, Kobayashi K, Kanno I, Suzuki N.
  • Organizer: XIII International Symposium on Blood Substitutes and Oxygen Therapeutics
  • Place of Presentation: Boston, USA
  • Year and Date: 2011-07-29
• [Presentation] The effect of Artificial RBCS injection for hemorrhagic hypotension in mice (2011)
  • Author(s): Tomita Y, Unekawa M, Toriumi H, Masamoto K, Sakai H, Tsuchida E, Horinouchi H, Kobayashi K, Kanno I, Suzuki N
  • Organizer: Brain 11 ; 25th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-28
• [Presentation] Prolonged moderate hypoxia impairs neuro-glia-vascular coupling in the somatosensory cortex accompanied by parechymal microvessel dilation and astroglia swelling in mice (2011)
  • Author(s): Kanno I, Masamoto K, Takuwa H, Kawaguchi H, Tomita Y, Suzuki N, Obata T
  • Organizer: Brain 11 ; 25th International Symposium on Cerebr al Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-26
• [Presentation] Cerebral blood flow restoration mechanisms after unilateral common carotid artery occlusion in mice (2011)
  • Author(s): Yamada S, Guo H, Itoh Y, Toriumi H, Tomita Y, Hoshino H, Suzuki N
  • Organizer: Brain 11 ; 25th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-26
• [Presentation] Resident endothelial cells surrounding damaged arterial endothelium reend othelialize the lesion without any involvement of foreign progenitor cells (2011)
  • Author(s): Itoh Y, Toriumi H, Yamada S, Tomita Y, Hoshino H, Suzuki N
  • Organizer: Brain 11 ; 25th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-26
• [Presentation] Transhemispheric effect of cortical spreading depression : electroencephalogram was suppressed but erythrocyte velocity in intraparenchymal capillaries was not affected in rat (2011)
  • Author(s): Unekawa M, Tomita Y, Toriumi H, Masamoto K, Kanno I, Suzuki N
  • Organizer: Brain 11 ; 25th International Symposium on Cerebral Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-26
• [Presentation] Longitudinal imaging of glio-vascular remodeling induced by chronic hypoxia in mouse somatosensory cortex (2011)
  • Author(s): Masamoto K, Tomita Y, Takuwa H, Unekawa M, Toriumi H, Itoh Y, Suzuki N., Kanno I
  • Organizer: Brain 11 ; 25th International Symposium on Cerebr al Blood Flow, Metabolism & Function
  • Place of Presentation: Barcelona
  • Year and Date: 2011-05-24