Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Restorative effects of human neural stem cell grafts on the primate spinal cord

Abstract

We grafted human spinal cord–derived neural progenitor cells (NPCs) into sites of cervical spinal cord injury in rhesus monkeys (Macaca mulatta). Under three-drug immunosuppression, grafts survived at least 9 months postinjury and expressed both neuronal and glial markers. Monkey axons regenerated into grafts and formed synapses. Hundreds of thousands of human axons extended out from grafts through monkey white matter and synapsed in distal gray matter. Grafts gradually matured over 9 months and improved forelimb function beginning several months after grafting. These findings in a 'preclinical trial' support translation of NPC graft therapy to humans with the objective of reconstituting both a neuronal and glial milieu in the site of spinal cord injury.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Graft concept, procedure, survival, and differentiation.
Figure 2: Changes in graft density and cell size.
Figure 3: Axon emergence from grafts.
Figure 4: Host axons regenerate into human NPC grafts.

References

  1. 1

    Blesch, A. & Tuszynski, M.H. Spinal cord injury: plasticity, regeneration and the challenge of translational drug development. Trends Neurosci. 32, 41–47 (2009).

    Article  CAS  PubMed  Google Scholar 

  2. 2

    Fawcett, J.W. Overcoming inhibition in the damaged spinal cord. J. Neurotrauma 23, 371–383 (2006).

    Article  PubMed  Google Scholar 

  3. 3

    Fitch, M.T. & Silver, J. CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. Exp. Neurol. 209, 294–301 (2008).

    Article  CAS  PubMed  Google Scholar 

  4. 4

    He, Z. & Koprivica, V. The Nogo signaling pathway for regeneration block. Annu. Rev. Neurosci. 27, 341–368 (2004).

    Article  CAS  PubMed  Google Scholar 

  5. 5

    Schwab, M.E. Nogo and axon regeneration. Curr. Opin. Neurobiol. 14, 118–124 (2004).

    Article  CAS  PubMed  Google Scholar 

  6. 6

    Lu, P. & Tuszynski, M.H. Growth factors and combinatorial therapies for CNS regeneration. Exp. Neurol. 209, 313–320 (2008).

    Article  CAS  PubMed  Google Scholar 

  7. 7

    Filbin, M.T. Recapitulate development to promote axonal regeneration: good or bad approach? Phil. Trans. R. Soc. Lond. B 361, 1565–1574 (2006).

    Article  CAS  Google Scholar 

  8. 8

    Kadoya, K. et al. Combined intrinsic and extrinsic neuronal mechanisms facilitate bridging axonal regeneration one year after spinal cord injury. Neuron 64, 165–172 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. 9

    Jakeman, L.B. & Reier, P.J. Axonal projections between fetal spinal cord transplants and the adult rat spinal cord: a neuroanatomical tracing study of local interactions. J. Comp. Neurol. 307, 311–334 (1991).

    Article  CAS  PubMed  Google Scholar 

  10. 10

    Reier, P.J., Stokes, B.T., Thompson, F.J. & Anderson, D.K. Fetal cell grafts into resection and contusion/compression injuries of the rat and cat spinal cord. Exp. Neurol. 115, 177–188 (1992).

    Article  CAS  PubMed  Google Scholar 

  11. 11

    Wictorin, K. & Björklund, A. Axon outgrowth from grafts of human embryonic spinal cord in the lesioned adult rat spinal cord. Neuroreport 3, 1045–1048 (1992).

    Article  CAS  PubMed  Google Scholar 

  12. 12

    Coumans, J.V. et al. Axonal regeneration and functional recovery after complete spinal cord transection in rats by delayed treatment with transplants and neurotrophins. J. Neurosci. 21, 9334–9344 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. 13

    Cummings, B.J. et al. Human neural stem cells differentiate and promote locomotor recovery in spinal cord–injured mice. Proc. Natl. Acad. Sci. USA 102, 14069–14074 (2005).

    Article  CAS  PubMed  Google Scholar 

  14. 14

    Salazar, D.L., Uchida, N., Hamers, F.P.T., Cummings, B.J. & Anderson, A.J. Human neural stem cells differentiate and promote locomotor recovery in an early chronic spinal cord injury NOD-scid mouse model. PLoS One 5, e12272 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. 15

    Bonner, J.F. et al. Grafted neural progenitors integrate and restore synaptic connectivity across the injured spinal cord. J. Neurosci. 31, 4675–4686 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. 16

    Courtine, G. et al. Recovery of supraspinal control of stepping via indirect propriospinal relay connections after spinal cord injury. Nat. Med. 14, 69–74 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. 17

    Lu, P. et al. Long-distance growth and connectivity of neural stem cells after severe spinal cord injury. Cell 150, 1264–1273 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. 18

    Lu, P. et al. Long-distance axonal growth from human induced pluripotent stem cells after spinal cord injury. Neuron 83, 789–796 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. 19

    Kadoya, K. et al. Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration. Nat. Med. 22, 479–487 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. 20

    Rosenzweig, E.S. et al. Extensive spinal decussation and bilateral termination of cervical corticospinal projections in rhesus monkeys. J. Comp. Neurol. 513, 151–163 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  21. 21

    Kuypers, H.G.J.M. in Handbook of Physiology (eds. Brooks, V.B., Brookhart, J.M. & Mountcastle, V.B.). Anatomy of the descending pathways (The American Physiological Society, 1981).

  22. 22

    Lacroix, S. et al. Bilateral corticospinal projections arise from each motor cortex in the macaque monkey: a quantitative study. J. Comp. Neurol. 473, 147–161 (2004).

    Article  PubMed  Google Scholar 

  23. 23

    Galea, M.P. & Darian-Smith, I. Manual dexterity and corticospinal connectivity following unilateral section of the cervical spinal cord in the macaque monkey. J. Comp. Neurol. 381, 307–319 (1997).

    Article  CAS  PubMed  Google Scholar 

  24. 24

    Kwon, B.K. et al. Large animal and primate models of spinal cord injury for the testing of novel therapies. Exp. Neurol. 269, 154–168 (2015).

    Article  PubMed  Google Scholar 

  25. 25

    Tuszynski, M.H. in Translational Neuroscience 1st edn. (ed. Tuszynski, M.H.) Conclusion (Springer US, 2016).

  26. 26

    Anderson, A.J., Piltti, K.M., Hooshmand, M.J., Nishi, R.A. & Cummings, B.J. Preclinical efficacy failure of human CNS-derived stem cells for use in the pathway study of cervical spinal cord injury. Stem Cell Rep. 8, 249–263 (2017).

    Article  Google Scholar 

  27. 27

    Rosenzweig, E.S. et al. Extensive spontaneous plasticity of corticospinal projections after primate spinal cord injury. Nat. Neurosci. 13, 1505–1510 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. 28

    Cizkova, D. et al. Functional recovery in rats with ischemic paraplegia after spinal grafting of human spinal stem cells. Neuroscience 147, 546–560 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. 29

    Guo, X., Johe, K., Molnar, P., Davis, H. & Hickman, J. Characterization of a human fetal spinal cord stem cell line, NSI-566RSC, and its induction to functional motoneurons. J. Tissue Eng. Regen. Med. 4, 181–193 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. 30

    Glass, J.D. et al. Lumbar intraspinal injection of neural stem cells in patients with amyotrophic lateral sclerosis: results of a phase I trial in 12 patients. Stem Cells 30, 1144–1151 (2012).

    Article  CAS  PubMed  Google Scholar 

  31. 31

    Sun, W. et al. SOX9 Is an astrocyte-specific nuclear marker in the adult brain outside the neurogenic regions. J. Neurosci. 37, 4493–4507 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. 32

    Ren, Y. et al. Ependymal cell contribution to scar formation after spinal cord injury is minimal, local and dependent on direct ependymal injury. Sci. Rep. 7, 41122 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. 33

    Golan, N. et al. Identification of Tmem10/Opalin as an oligodendrocyte enriched gene using expression profiling combined with genetic cell ablation. Glia 56, 1176–1186 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  34. 34

    Lu, P. et al. Prolonged human neural stem cell maturation supports recovery in injured rodent CNS. J. Clin. Invest. 127, 3287–3299 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  35. 35

    Custo Greig, L.F., Woodworth, M.B., Galazo, M.J., Padmanabhan, H. & Macklis, J.D. Molecular logic of neocortical projection neuron specification, development and diversity. Nat. Rev. Neurosci. 14, 755–769 (2013).

    Article  CAS  Google Scholar 

  36. 36

    Fields, H.L., Heinricher, M.M. & Mason, P. Neurotransmitters in nociceptive modulatory circuits. Annu. Rev. Neurosci. 14, 219–245 (1991).

    Article  CAS  PubMed  Google Scholar 

  37. 37

    Schmidt, B.J. & Jordan, L.M. The role of serotonin in reflex modulation and locomotor rhythm production in the mammalian spinal cord. Brain Res. Bull. 53, 689–710 (2000).

    Article  CAS  PubMed  Google Scholar 

  38. 38

    Nout, Y.S. et al. Methods for functional assessment after C7 spinal cord hemisection in the rhesus monkey. Neurorehabil. Neural Repair 26, 556–569 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  39. 39

    Nout, Y.S. et al. Animal models of neurologic disorders: a nonhuman primate model of spinal cord injury. Neurotherapeutics 9, 380–392 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. 40

    Salegio, E.A. et al. A unilateral cervical spinal cord contusion injury model in non-human primates (Macaca mulatta). J. Neurotrauma 33, 439–459 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  41. 41

    Friedli, L. et al. Pronounced species divergence in corticospinal tract reorganization and functional recovery after lateralized spinal cord injury favors primates. Sci. Transl. Med. 7, 302ra134 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  42. 42

    Windrem, M.S. et al. Fetal and adult human oligodendrocyte progenitor cell isolates myelinate the congenitally dysmyelinated brain. Nat. Med. 10, 93–97 (2004).

    Article  CAS  PubMed  Google Scholar 

  43. 43

    Osorio, M.J. & Goldman, S.A. Glial progenitor cell-based treatment of the childhood leukodystrophies. Exp. Neurol. 283 Part B, 476–488 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. 44

    Steward, O., Sharp, K.G., Yee, K.M., Hatch, M.N. & Bonner, J.F. Characterization of ectopic colonies that form in widespread areas of the nervous system with neural stem cell transplants into the site of a severe spinal cord injury. J. Neurosci. 34, 14013–14021 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. 45

    Tuszynski, M.H. et al. Neural stem cell dissemination after grafting to CNS injury sites. Cell 156, 388–389 (2014).

    Article  CAS  PubMed  Google Scholar 

  46. 46

    Mongiat, L.A., Espósito, M.S., Lombardi, G. & Schinder, A.F. Reliable activation of immature neurons in the adult hippocampus. PLoS One 4, e5320 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. 47

    Dieni, C.V. et al. Low excitatory innervation balances high intrinsic excitability of immature dentate neurons. Nat. Commun. 7, 11313 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. 48

    Hollis, E.R. II., Jamshidi, P., Löw, K., Blesch, A. & Tuszynski, M.H. Induction of corticospinal regeneration by lentiviral trkB-induced Erk activation. Proc. Natl. Acad. Sci. USA 106, 7215–7220 (2009).

    Article  PubMed  Google Scholar 

  49. 49

    Ghosh, M. et al. Extensive cell migration, axon regeneration, and improved function with polysialic acid-modified Schwann cells after spinal cord injury. Glia 60, 979–992 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  50. 50

    Liu, K. et al. PTEN deletion enhances the regenerative ability of adult corticospinal neurons. Nat. Neurosci. 13, 1075–1081 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. 51

    Jin, D. et al. Restoration of skilled locomotion by sprouting corticospinal axons induced by co-deletion of PTEN and SOCS3. Nat. Commun. 6, 8074 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. 52

    Tuszynski, M.H., Gabriel, K., Gerhardt, K. & Szollar, S. Human spinal cord retains substantial structural mass in chronic stages after injury. J. Neurotrauma 16, 523–531 (1999).

    Article  CAS  PubMed  Google Scholar 

  53. 53

    Lu, P., Ahmad, R. & Tuszynski, M.H. in Translational Neuroscience 1st edn. (ed. Tuszynski, M.H.) Neural Stem Cells for Spinal Cord Injury (Springer US, 2016).

  54. 54

    Christie, K.J., Webber, C.A., Martinez, J.A., Singh, B. & Zochodne, D.W. PTEN inhibition to facilitate intrinsic regenerative outgrowth of adult peripheral axons. J. Neurosci. 30, 9306–9315 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. 55

    Schmid, A.C., Byrne, R.D., Vilar, R. & Woscholski, R. Bisperoxovanadium compounds are potent PTEN inhibitors. FEBS Lett. 566, 35–38 (2004).

    Article  CAS  PubMed  Google Scholar 

  56. 56

    Raff, M.C. et al. Programmed cell death and the control of cell survival: lessons from the nervous system. Science 262, 695–700 (1993).

    Article  CAS  PubMed  Google Scholar 

  57. 57

    Raff, M.C. Size control: the regulation of cell numbers in animal development. Cell 86, 173–175 (1996).

    Article  CAS  PubMed  Google Scholar 

  58. 58

    Jacobson, M.D., Weil, M. & Raff, M.C. Programmed cell death in animal development. Cell 88, 347–354 (1997).

    Article  CAS  PubMed  Google Scholar 

  59. 59

    Maor-Nof, M. & Yaron, A. Neurite pruning and neuronal cell death: spatial regulation of shared destruction programs. Curr. Opin. Neurobiol. 23, 990–996 (2013).

    Article  CAS  PubMed  Google Scholar 

  60. 60

    Kaiser, H.F. Directional statistical decisions. Psychol. Rev. 67, 160–167 (1960).

    Article  CAS  PubMed  Google Scholar 

  61. 61

    Cattell, R.B. The scree test for the number of Factors. Multivariate Behav. Res. 1, 245–276 (1966).

    Article  CAS  PubMed  Google Scholar 

  62. 62

    Guadagnoli, E. & Velicer, W.F. Relation of sample size to the stability of component patterns. Psychol. Bull. 103, 265–275 (1988).

    Article  PubMed  Google Scholar 

  63. 63

    Ferguson, A.R. et al. Derivation of multivariate syndromic outcome metrics for consistent testing across multiple models of cervical spinal cord injury in rats. PLoS One 8, e59712 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. 64

    Little, R.J.A. & Rubin, D.B. Statistical Analysis with Missing Data 2nd edn (John Wiley & Sons, Inc., 2002).

  65. 65

    Bacchetti, P., Deeks, S.G. & McCune, J.M. Breaking free of sample size dogma to perform innovative translational research. Sci. Transl. Med. 3, 87ps24 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Human 566RSC-UBQT neural stem cells were a gift from NeuralStem, Inc. This work was supported by the Veterans Administration (Gordon Mansfield Spinal Cord Injury Collaborative Consortium, RR&D B7332R, MHT; RR&D RX001045, JHB), the National Institutes of Health (R01 NS042291, MHT; R01 NS104442, MHT), the Department of Defense (W81XWH-12-1-0592; E.S.R.), the Craig H. Neilsen Foundation (M.H.T.), the Bernard and Anne Spitzer Charitable Trust (M.H.T.), and the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation (M.H.T.).

Author information

Affiliations

Authors

Contributions

E.S.R., J.H.B., P.L., E.A.S., K.K., L.A.H., Y.S.N.-L., A.R.F., M.S.B., J.C.B., and M.H.T. designed the experiments. E.S.R., J.H.B., P.L., H.K., E.A.S., K.K., R.M., S.H., Y.S.N.-L., J.C.B., and M.H.T. carried out the experiments. E.S.R., J.H.B., H.K., E.A.S., J.L.W., J.J.L., J.R.H., L.A.H., A.R.F., and M.H.T. analyzed the data. E.S.R. and M.H.T. wrote the manuscript. E.S.R., J.H.B., P.L., H.K., E.A.S., K.K., J.L.W., J.J.L., R.M., S.H., J.R.H., L.A.H., Y.S.N.-L., A.R.F., M.S.B., J.C.B., and M.H.T. edited the manuscript.

Corresponding author

Correspondence to Mark H Tuszynski.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

Supplementary Figures 1–15 (PDF 15811 kb)

Life Sciences Reporting Summary (PDF 180 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Rosenzweig, E., Brock, J., Lu, P. et al. Restorative effects of human neural stem cell grafts on the primate spinal cord. Nat Med 24, 484–490 (2018). https://doi.org/10.1038/nm.4502

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing