Neural Stem Cell Transplantation into a Mouse Model of Stroke

  • Alejandro De Los AngelesEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2005)


Stroke is the fifth leading cause of death among Americans each year. Current standard-of-care treatment for stroke deploys intravenous tissue-type plasminogen activator (tPA), mechanical thrombolysis, or delivery of fibrinolytics. Although these therapies have reduced stroke-induced damage, therapeutic options still remain limited. Transplantation of patient-specific neural stem (NS) cells represents a promising strategy for the treatment of stroke. Basic science research has shown that transplanted NS cells can differentiate in the brain of rodent models of stroke and promote behavioral recovery. Clinical trials exploring the feasibility of stem cell treatment for stroke are currently being conducted. However, questions remain regarding the optimal means of delivering NS cells, including cell dose, infusion speed, timing of transplantation, anatomic site, and imaging-assisted monitoring and guidance. Of the different available delivery modalities, intravascular NS delivery after stroke represents one practical approach. In this chapter, I provide methods for intravascular delivery of NS cells in a mouse model of stroke. The techniques involved include cell culture of NS cells, flow cytometry of NS cells, modeling stroke via unilateral common carotid artery occlusion, intra-arterial injection of NS cells into the brain, behavior analyses, and immunohistochemistry. Intra-arterial NS cell therapy has the potential to improve functional recovery after ischemic stroke.

Key words

Stroke VLA4 CD49d CCR2 VCAM1 CCL2 Postnatal chimera Neural stem cell Common carotid artery Hypoxia-ischemia Mouse stroke model C17.2 neural progenitors 


  1. 1.
    CDC Stroke facts. 2017.
  2. 2.
    Sinden JD, Hicks C, Stroemer P, Vishnubhatla I, Corteling R (2017) Human neural stem cell therapy for chronic ischemic stroke: charting progress from laboratory to patients. Stem Cells Dev 26:933–947CrossRefGoogle Scholar
  3. 3.
    Kelly S, Bliss TM, Shah AK, Sun GH, Ma M, Foo WC, Masel J, Yenari MA, Weissman IL, Uchida N, Palmer T, Steinberg GK (2004) Transplanted human fetal neural stem cells survive, migrate, and differentiate in ischemic rat cerebral cortex. PNAS 101:11839–11844CrossRefGoogle Scholar
  4. 4.
    Guzman R, Uchida N, Bliss TM, He D, Christopherson KK, Stellwagen D, Capela A, Greve J, Malenka RC, Moseley ME, Palmer TD, Steinberg GK (2007) Long-term monitoring of transplanted human neural stem cells in developmental and pathological contexts with MRI. PNAS 104:10211–10216CrossRefGoogle Scholar
  5. 5.
    Guzman R, Janowski M, Walczak P (2018) Intra-arterial delivery of cell therapies for stroke. Stroke 49:1075–1082CrossRefGoogle Scholar
  6. 6.
    Pluchino S, Zanotti L, Rossi B, Brambilla E, Ottoboni L, Salani G, Martinello M, Cattalini A, Bergami A, Furlan R, Comi G, Constantin G, Martino G (2005) Neurosphere-derived multipotent precursors promote neuroprotection by an immunomodulatory mechanism. Nature 436:266–271CrossRefGoogle Scholar
  7. 7.
    Andres RH, Choi R, Pendharkar AV, Gaeta X, Wang N, Nathan JK, Chua JY, Lee SW, Palmer TD, Steinberg GK, Guzman R (2011) The CCR2/CCL2 interaction mediates the transendothelial recruitment of intravascularly delivered neural stem cells to the ischemic brain. Stroke 42:2923–2931CrossRefGoogle Scholar
  8. 8.
    Frijns CJ, Kappelle LJ (2002) Inflammatory cell adhesion molecules in ischemic cerebrovascular disease. Stroke 33:2115–2122CrossRefGoogle Scholar
  9. 9.
    Pluchino S, Quattrini A, Brambilla E, Gritti A, Salani G, Dina G, Galli R, Del Carro U, Amadio S, Bergami A, Furlan R, Comi G, Vescovi AL, Martino G (2003) Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis. Nature 422:688–694CrossRefGoogle Scholar
  10. 10.
    Mueller FJ, Sereobyan N, Schraufstatter IU, DiScipio R, Waeman D, Loring JF et al (2006) Adhesive interactions between human neural stem cells and inflamed human vascular endothelium are mediated by integrins. Stem Cells 24:2367–2372CrossRefGoogle Scholar
  11. 11.
    Rebenko-Moll NM, Liu L, Cardona A, Ransohoff RM (2006) Chemokines, mononuclear cells and the nervous system: heaven (or hell) is in the details. Curr Opin Immunol 18:683–689CrossRefGoogle Scholar
  12. 12.
    Tran PB, Ren D, Veldhouse TJ, Miller RJ (2004) Chemokine receptors are expressed widely by embryonic and adult neural progenitor cells. J Neurosci Res 76:20–34CrossRefGoogle Scholar
  13. 13.
    Belmadani A, Tran PB, Ren D, Miller RJ (2006) Chemokines regulate the migration of neural progenitors to sites of neuroinflammation. J Neurosci 26:3182–3191CrossRefGoogle Scholar
  14. 14.
    Robin AM, Zhang ZG, Wang L, Zhang RL, Katakowski M, Zhang L et al (2006) Stromal cell-derived factor 1 alpha mediates neural progenitor cell motility after focal cerebral ischemia. J Cereb Blood Flow Metab 26:125–134CrossRefGoogle Scholar
  15. 15.
    Yan YP, Sailor KA, Lang BT, Park SW, Vemuganti R, Dempsey RJ (2007) Monocyte chemoattractant protein-1 plays a critical role in neuroblast migration after focal cerebral ischemia. J Cereb Blood Flow Metab 27:1213–1224CrossRefGoogle Scholar
  16. 16.
    Friedrich MA, Martins MP, Araujo MD, Klamt C, Vedolin L, Garichochea B et al (2012) Intra-arterial infusion of autologous bone marrow mononuclear cells in patients with moderate to severe middle cerebral artery acute ischemic stroke. Cell Transplant 21(Supp 1):S13–S21CrossRefGoogle Scholar
  17. 17.
    Snyder EY, Deitcher DL, Walsh C, Arnold-Aldea S, Hartwieg EA, Cepko CL (1992) Multipotent neural cell lines can engraft and participate in development of mouse cerebellum. Cell 68:33–51CrossRefGoogle Scholar
  18. 18.
    Snyder EY, Taylor RM, Wolfe JH (1995) Neural progenitor cell engraftment corrects lysosomal storage throughout the MPS VII mouse brain. Nature 374:367–370CrossRefGoogle Scholar
  19. 19.
    Ryder EF, Snyder EY, Cepko CL (1990) Establishment and characterization of multipotent neural cell lines using retrovirus vector-mediated oncogene transfer. J Neurobiol 21:356–375CrossRefGoogle Scholar
  20. 20.
    Guzman R, De Los AA, Cheshier S, Choi R, Hoang S, Liauw J, Schaar B, Steinberg GK (2008) Intracarotid injection of fluorescence activated cell-sorted CD49d-positive neural stem cells improves target cell delivery and behavior after stroke in a mouse stroke model. Stroke 39:1300–1306CrossRefGoogle Scholar
  21. 21.
    Engel O, Kolodzij S, Dirnagl U, Prinz V (2011) Modeling stroke in mice—middle cerebral artery occlusion with the filament model. J Vis Exp 6:47Google Scholar
  22. 22.
    Rosenblum S, Wang N, Smith TN, Pendharkar AV, Chua JY, Birk H, Guzman R (2012) Timing of intra-arterial neural stem cell transplantation after hypoxia-ischemia influences cell engraftment, survival and differentiation. Stroke 43:1624–1631CrossRefGoogle Scholar
  23. 23.
    Deacon RMJ (2013) Measuring motor coordination in mice. J Vis Exp 75:2609Google Scholar
  24. 24.
    Gage GJ, Kipke DR, Shain W (2012) Whole animal perfusion fixation for rodents. J Vis Exp 65:3564Google Scholar
  25. 25.
    Magavi SS, Macklis JD (2008) Identification of newborn cells by BrdU labeling and immunocytochemistry in vivo. Methods Mol Biol 438:335–343CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of PsychiatryYale University School of MedicineNew HavenUSA

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