Neurotoxicity Research

, Volume 7, Issue 1–2, pp 169–177 | Cite as

Treatment of spinal cord injury with co-grafts of genetically modified schwann cells and fetal spinal cord cell suspension in the rat

  • Shi -Qing Feng
  • Xiao -Hong Kong
  • Shi -Fu Guo
  • Pei Wang
  • Li Li
  • Jin -Hua Zhong
  • Xin -Fu Zhou


Epub ahead of print: December 2004 Fetal spinal cord cells, Schwann cells and neurotrophins all have the capacity to promote repair of injured spinal cord in animal models. To explore the possibility of using these approaches to treat clinical patients, we have examined whether a combination of these protocols produces functional and anatomical improvement. The spinal cords of adult rats (n=16) were injured with a modified New York University (NYU) device (10 gram.5cm). One week after injury, the injured cords were injected with Dulbecco-modified Eagles Medium (DMEM, control group), or fetal spinal cord cell suspension (FSCS) plus nerve growth factor (NGF) gene-modified Schwann cells (SC) and brain-derived neurotrophic factor (BDNF) genemodified SC (treatment group). The rats were subjected to BBB (Basso, Beattie, Bresnahan, Exp. Neurol. 139:244, 1996) behavioral tests. Anterograde tracing of corticospinal tract was performed before sacrifice 3 months after the treatment. The results showed that the combination treatment elicited a robust growth of corticospinal axons within and beyond the injury site. A dramatic functional recovery in the treatment group was observed compared with the control group. We conclude that the combination of FSCS with genetically modified Schwann cells over-expressing NGF and BDNF was an effective protocol for the treatment of severe spinal cord injury.


Schwann cell Fetal spinal cord cells Regeneration Nerve growth factor Brain-derived neurotrophic factor Corticospinal tract Behavior 


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  1. Akesson E, A Kjaeldgaard and A Seiger (1998) Human embryonic spinal cord grafts in adult rat spinal cord cavities: survival, growth, and interactions with the host.Exp. Neurol. 149, 262–276.PubMedCrossRefGoogle Scholar
  2. Anderson DK, DR Howland and PJ Reier (1995) Fetal neural grafts and repair of the injured spinal cord.Brain Pathol. 5, 451–457.PubMedCrossRefGoogle Scholar
  3. Andrade-Rozental AF, R Rozental, A Hassankhani, DC Spray and HJ Federoff (1995) Characterization of two populations of ectopic cells isolated from the hearts of NGF transgenic mice.Dev. Biol. 169, 533–546.PubMedCrossRefGoogle Scholar
  4. Bamber NI, H Li, P Aebischer and XM Xu (1999) Fetal spinal cord tissue in mini-guidance channels promotes longitudinal axonal growth after grafting into hemisected adult rat spinal cords.NeuralPlast. 6, 103–121.Google Scholar
  5. Bartolomei JC and CA Greer (2000) Olfactory ensheathing cells: bridging the gap in spinal cord injury.Neurosurgery 47, 1057- 1069.PubMedCrossRefGoogle Scholar
  6. Basso DM, MS Beattie and JC Bresnahan (1996) Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection.Exp. Neurol. 139, 244–256.PubMedCrossRefGoogle Scholar
  7. Bernstein-Goral H and BS Bregman (1997) Axotomized rubrospinal neurons rescued by fetal spinal cord transplants maintain axon collaterals to rostral CNS targets.Exp. Neurol. 148, 13–25.PubMedCrossRefGoogle Scholar
  8. Bernstein-Goral H, PS Diener and BS Bregman (1997) Regenerating and sprouting axons differ in their requirements for growth after injury.Exp. Neurol. 148, 51–72.PubMedCrossRefGoogle Scholar
  9. Blesch A and MH Tuszynski (1997) Robust growth of chronically injured spinal cord axons induced by grafts of genetically modified NGF-secreting cells.Exp. Neurol. 148, 444–452.PubMedCrossRefGoogle Scholar
  10. Bradbury EJ, S Khemani, R Von, VR King, JV Priestley and SB McMahon (1999) NT-3 promotes growth of lesioned adult rat sensory axons ascending in the dorsal columns of the spinal cord.Eur. J. Neurosci. 11, 3873–3883.PubMedCrossRefGoogle Scholar
  11. Bregman BS (1998) Regeneration in the spinal cord.Curr. Opin. Neurobiol. 8, 800–807.PubMedCrossRefGoogle Scholar
  12. Bregman BS, M McAtee, HN Dai and PL Kuhn (1997) Neurotrophic factors increase axonal growth after spinal cord injury and transplantation in the adult rat.Exp. Neurol. 148, 475- 494.PubMedCrossRefGoogle Scholar
  13. Bregman BS, E Broude, M McAtee and MS Kelley (1998) Transplants and neurotrophic factors prevent atrophy of mature CNS neurons after spinal cord injury.Exp. Neurol. 149, 13–27.PubMedCrossRefGoogle Scholar
  14. Chan JR, JM Cosgaya, TJ Wu and EM Shooter (2001) Neurotrophins are key mediators of the myelination program in the peripheral nervous system.Proc. Natl. Acad. Sci. USA 98, 14661–14668.PubMedCrossRefGoogle Scholar
  15. Constantini S and W Young (1994) The effects of methylpred- nisolone and the ganglioside GM1 on acute spinal cord injury in rats.J. Neurosurgery 80, 97–111.Google Scholar
  16. Coumans JV, TT Lin, HN Dai, L MacArthur, M McAtee, C Nash and BS Bregman (2001) Axonal regeneration and functional recovery after complete spinal cord transection in rats by delayed treatment with transplants and neurotrophins.J. Neurosci. 21, 9334–9344.PubMedGoogle Scholar
  17. Day-Lollini PA, GR Stewart, MJ Taylor, RM Johnson and GJ Chellman (1997) Hyperplastic changes within the leptomeninges of the rat and monkey in response to chronic intracerebroventricular infusion of nerve growth factor.Exp. Neurol. 145, 24–37.PubMedCrossRefGoogle Scholar
  18. Diener PS and BS Bregman (1998) Fetal spinal cord transplants support the development of target reaching and coordinated postural adjustments after neonatal cervical spinal cord injury.J. Neurosci. 18, 763–778.PubMedGoogle Scholar
  19. Duchossoy Y, L Kassar-Duchossoy, D Orsal, O Stettler and JC Horvat (2001) Reinnervation of the biceps brachii muscle following cotransplantation of fetal spinal cord and autologous peripheral nerve into the injured cervical spinal cord of the adult rat.Exp. Neurol. 167, 329–340.PubMedCrossRefGoogle Scholar
  20. Fawcett JW (1998) Spinal cord repair: from experimental models to human application.Spinal Cord 36, 811–817.PubMedCrossRefGoogle Scholar
  21. Feng SQ, SF Guo, JT Chen, JC Chen, XH Kong, W P, and XL Ma (2000) A study on the NGF and BDNF genetically modified Schwann cells. Chin.J. Orthopaedics 20, 488–492.Google Scholar
  22. Ferguson IA, T Koide and RA Rush (2001) Stimulation of corti- cospinal tract regeneration in the chronically injured spinal cord.Eur. J. Neurosci. 13, 1059–1064.PubMedCrossRefGoogle Scholar
  23. Frostick SP, Q Yin and GJ Kemp (1998) Schwann cells, neurotrophic factors, and peripheral nerve regeneration.Microsurgery 18, 397–405.PubMedCrossRefGoogle Scholar
  24. Fu SY and T Gordon (1997) The cellular and molecular basis of peripheral nerve regeneration.Mol. Neurobiol. 14, 67–116.PubMedCrossRefGoogle Scholar
  25. Funakoshi H, J Frisén, G Barbany, T Timmusk, O Zachrisson, VMK Verge and H Persson (1993) Differential expression of mRNAs for neurotrophins and their receptors after axotomy of the sciatic nerve.J. Cell Biol. 123, 455–465.PubMedCrossRefGoogle Scholar
  26. Guest JD, A Rao, L Olson, MB Bunge and RP Bunge (1997) The ability of human Schwann cell grafts to promote regeneration in the transected nude rat spinal cord.Exp. Neurol. 148, 502–522.PubMedCrossRefGoogle Scholar
  27. Hagg T, M Rende, E Magal, P Burnham, M Oudega and S Varon (1993) Potential regulation by trophic factors of low-affinity NGF receptors in spinal motor neurons.Brain Res. Bull. 30, 347–352.PubMedCrossRefGoogle Scholar
  28. Ikeda O, M Murakami, H Ino, M Yamazaki, M Koda, C Nakayama and H Moriya (2002) Effects of brain-derived neurotrophic factor (BDNF) on compression- induced spinal cord injury: BDNF attenuates down-regulation of superoxide dismutase expression and promotes up-regulation of myelin basic protein expression.J. Neuropathol. Exp. Neurol. 61, 142–153.PubMedGoogle Scholar
  29. Iwashita Y, S Kawaguchi and M Murata (1994) Restoration of function by replacement of spinal cord segments in the rat.Nature 367, 167–170.PubMedCrossRefGoogle Scholar
  30. Jirsova K, P Sodaar, V Mandys and PR Bar (1997) Cold jet: a method to obtain pure Schwann cell cultures without the need for cytotoxic, apoptosis-inducing drug treatment.J. Neurosci. Methods 78, 133–137.PubMedCrossRefGoogle Scholar
  31. Keirstead HS, SV Morgan, MJ Wilby and JW Fawcett (1999) Enhanced axonal regeneration following combined demyelination plus Schwann cell transplantation therapy in the injured adult spinal cord.Exp. Neurol. 159, 225–236.PubMedCrossRefGoogle Scholar
  32. Khan T, B Green and JR Perez-Polo (1987) Effect of injury on nerve growth factor uptake by sensory ganglia.J. Neurosci. Res. 18, 562–567.PubMedCrossRefGoogle Scholar
  33. Kim DH, PH Gutin, LJ Noble, D Nathan, JS Yu and RP Nockels (1996) Treatment with genetically engineered fibroblasts producing NGF or BDNF can accelerate recovery from traumatic spinal cord injury in the adult rat.NeuroReport 7, 2221–2225.PubMedCrossRefGoogle Scholar
  34. Kuhlengel KR, MB Bunge and RP Bunge (1990) Implantation of cultured sensory neurons and Schwann cells into lesioned neonatal rat spinal cord. I. Methods for preparing implants from dissociated cells.J. Comp. Neurol. 293, 63–73.PubMedCrossRefGoogle Scholar
  35. Kwon BK and W Tetzlaff (2001) Spinal cord regeneration: from gene to transplants.Spine 26, S13-S22.PubMedCrossRefGoogle Scholar
  36. Li Y, PM Field and G Raisman (1997) Repair of adult rat corticospinal tract by transplants of olfactory ensheathing cells.Science 277, 2000–2002.PubMedCrossRefGoogle Scholar
  37. Liang Y, JA Marcusson and O Johansson (1999) Light and electron microscopic immunohistochemical observations of p75 nerve growth factor receptor-immunoreactive dermal nerves in prurigo nodularis.Arch. Dermatol. Res. 291, 14–21.PubMedCrossRefGoogle Scholar
  38. Martin D, J Schoenen, P Delree, JM Rigo, B Rogister, P Leprince and G Moonen (1993) Syngeneic grafting of adult rat DRG- derived Schwann cells to the injured spinal cord.Brain Res. Bull. 30, 507–514.PubMedCrossRefGoogle Scholar
  39. Martin D, P Robe, R Franzen, P Delree, J Schoenen, A Stevenaert and G Moonen (1996) Effects of Schwann cell transplantation in a contusion model of rat spinal cord injury.J. Neurosci. Res. 45, 588–597.PubMedCrossRefGoogle Scholar
  40. McDonald JW, XZ Liu, Y Qu, S Liu, SK Mickey, D Turetsky, DI Gottlieb and DW Choi (1999) Transplanted embryonic stem cells survive, differentiate and promote recovery in injured rat spinal cord.Nat. Med. 5, 1410–1412.PubMedCrossRefGoogle Scholar
  41. Menei P, C Montero-Menei, SR Whittemore, RP Bunge and MB Bunge (1998) Schwann cells genetically modified to secrete human BDNF promote enhanced axonal regrowth across transected adult rat spinal cord.Eur. J. Neurosci. 10, 607–621.PubMedCrossRefGoogle Scholar
  42. Mizisin AP, M Bache, PS DiStefano, A Acheson, RM Lindsay and NA Calcutt (1997) BDNF attenuates functional and structural disorders in nerves of galactose-fed rats.J. Neuropathol. Exp. Neurol. 56, 1290–1301.PubMedCrossRefGoogle Scholar
  43. Murray M and I Fischer (2001) Transplantation and gene therapy: combined approaches for repair of spinal cord injury.Neuroscientist 7, 28–41.PubMedCrossRefGoogle Scholar
  44. Nicholls JG, H Vischer, Z Varga, S Erulkar and NR Saunders (1994) Repair of connections in injured neonatal and embryonic spinal cordin vitro. Prog. Brain Res. 103, 263–269.Google Scholar
  45. Oudega M, SE Gautier, P Chapon, M Fragoso, ML Bates, JM Parel and MB Bunge (2001) Axonal regeneration into Schwann cell grafts within resorbable poly(alpha-hydroxyacid) guidance channels in the adult rat spinal cord.Biomaterials 22, 1125–1136.PubMedCrossRefGoogle Scholar
  46. Paino CL, C Fernandez-Valle, ML Bates and MB Bunge (1994) Regrowth of axons in lesioned adult rat spinal cord, promotion by implants of cultured Schwann cells.J. Neurocytol. 23, 433- 452.PubMedCrossRefGoogle Scholar
  47. Pearse DD, FC Pereira, AE Marcillo, ML Bates, YA Berrocal, MT Filbin and MB Bunge (2004) cAMP and Schwann cells promote axonal growth and functional recovery after spinal cord injury.Nat. Med. 10, 610–616.PubMedCrossRefGoogle Scholar
  48. Plant GW, CL Christensen, M Oudega and MB Bunge (2003) Delayed transplantation of olfactory ensheathing glia promotes sparing/regeneration of supraspinal axons in the contused adult rat spinal cord.J. Neurotrauma 20, 1–16.PubMedCrossRefGoogle Scholar
  49. Raisman G (2001) Olfactory ensheathing cells — another miracle cure for spinal cord injury?Nat. Rev. Neurosci. 2, 369–375.PubMedCrossRefGoogle Scholar
  50. Ramon-Cueto A, GW Plant, J Avila and MB Bunge (1998) Longdistance axonal regeneration in the transected adult rat spinal cord is promoted by olfactory ensheathing glia transplants.J. Neurosci. 18, 3803–3815.PubMedGoogle Scholar
  51. Reier PJ, BT Stokes, FJ Thompson and DK Anderson (1992) Fetal cell grafts into resection and contusion/compression injuries of the rat and cat spinal cord.Exp. Neurol. 115, 177–188.PubMedCrossRefGoogle Scholar
  52. Ruitenberg MJ, GW Plant, FP Hamers, J Wortel, B Blits, PA Dijkhuizen, WH Gispen, GJ Boer and J Verhaagen (2003)Ex vivo adenoviral vector-mediated neurotrophin gene transfer to olfactory ensheathing glia: effects on rubrospinal tract regeneration, lesion size, and functional recovery after implantation in the injured rat spinal cord.J. Neurosci. 23, 7045–7058.PubMedGoogle Scholar
  53. Schecterson LC and M Bothwell (1992) Novel roles for neurotrophins are suggested by BDNF and NT-3 mRNA expression in developing neurons.Neuron 9, 449–463.PubMedCrossRefGoogle Scholar
  54. Schnell L, R Schneider, R Kolbeck, YA Barde and ME Schwab (1994) Neurotrophin-3 enhances sprouting of corticospinal tract during development and after adult spinal cord lesion.Nature 367, 170–173.PubMedCrossRefGoogle Scholar
  55. Schwartz M (2001) Immunological approaches to the treatment of spinal cord injury.Biodrugs 15, 585–593.PubMedCrossRefGoogle Scholar
  56. Schwartz M, O Lazarov-Spiegler, O Rapalino, I Agranov, G Velan and M Hadani (1999) Potential repair of rat spinal cord injuries using stimulated homologous macrophages.Neurosurgery 44, 1041–1045; discussion 1045–1046.PubMedCrossRefGoogle Scholar
  57. Skaper SD, M Manthorpe, R Adler and S Varon (1980) Survival, proliferation and morphological specialization of mouse Schwann cells in a serum-free, fully defined medium. J.Neurocytol. 9, 683–697.CrossRefGoogle Scholar
  58. Takami T, M Oudega, ML Bates, PM Wood, N Kleitman and MB Bunge (2002) Schwann cell but not olfactory ensheathing glia transplants improve hindlimb locomotor performance in the moderately contused adult rat thoracic spinal cord.J. Neurosci. 22, 6670–6681.PubMedGoogle Scholar
  59. Tuszynski MH, N Weidner, M McCormack, I Miller, H Powell and J Conner (1998) Grafts of genetically modified Schwann cells to the spinal cord, survival, axon growth, and myelination.J. Neuropathol. Exp. Neurol. 57, 205–217.CrossRefGoogle Scholar
  60. Weidner N, A Blesch, RJ Grill and MH Tuszynski (1999) Nerve growth factor-hypersecreting Schwann cell grafts augment and guide spinal cord axonal growth and remyelinate central nervous system axons in a phenotypically appropriate manner that correlates with expression of L1.J. Comp. Neurol. 413, 495–506.PubMedCrossRefGoogle Scholar
  61. Whitworth IH, RA Brown, CJ Dore, P Anand, CJ Green and G Terenghi (1996) Nerve growth factor enhances nerve regeneration through fibronectin grafts.J. Hand Surg. [Br.] 21, 514–522.Google Scholar
  62. Winkler J, GA Ramirez, HG Kuhn, DA Peterson, PA Day-Lollini, GR Stewart, MH Tuszynski, FH Gage and LJ Thal (1997) Reversible Schwann cell hyperplasia and sprouting of sensory and sympathetic neurites after intraventricular administration of nerve growth factor.Ann. Neurol. 41, 82–93.PubMedCrossRefGoogle Scholar
  63. Winkler J, GA Ramirez, LJ Thal and JJ Waite (2000) Nerve growth factor (NGF) augments cortical and hippocampal cholinergic functioning after p75NGF receptor-mediated deafferentation but impairs inhibitory avoidance and induces fear-related behaviors.J. Neurosci. 20, 834–844.PubMedGoogle Scholar
  64. Xu XM, V Guenard, N Kleitman, P Aebi scher and MB Bunge (1995) A combination of BDNF andNT-3 promotes supraspinal axonal regeneration into Schwann cell grafts in adult rat thoracic spinal cord.Exp. Neurol. 134, 261–272.PubMedCrossRefGoogle Scholar
  65. Xu XM, SX Zhang, H Li, P Aebischer and MB Bunge (1999) Regrowth of axons into the distal spinal cord through a Schwann- cell-seeded mini-channel implanted into hemisected adult rat spinal cord.Eur. J. Neurosci. 11, 1723–1740.PubMedCrossRefGoogle Scholar
  66. Ye JH and JD Houle (1997) Treatment of the chronically injured spinal cord with neurotrophic factors can promote axonal regeneration from supraspinal neurons.Exp. Neurol. 143, 70–81.PubMedCrossRefGoogle Scholar
  67. Yip HK, KM Rich, PA Lampe and EM Johnson Jr (1984) The effects of nerve growth factor and its antiserum on the postnatal development and survival after injury of sensory neurons in rat dorsal root ganglia.J. Neurosci. 4, 2986–2992.PubMedGoogle Scholar
  68. Zhang JY, XG Luo, CJ Xian, ZH Liu and X-F Zhou (2000) Endogenous BDNF is required for myelination and regeneration of injured sciatic nerve in rodents.Eur. J. Neurosci. 12, 4171- 4180.PubMedCrossRefGoogle Scholar
  69. Zhang Q, W Liao, Z Wang and Y Wu (2001) Effect of fetal spinal cord graft with different methods on axonal pathology after spinal cord contusion.Chin. J. Traumatol. 4, 147–151.PubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Shi -Qing Feng
    • 1
  • Xiao -Hong Kong
    • 2
  • Shi -Fu Guo
    • 1
  • Pei Wang
    • 1
  • Li Li
    • 3
  • Jin -Hua Zhong
    • 3
  • Xin -Fu Zhou
    • 3
  1. 1.Department of OrthopaedicTianjin Medical University HospitalTianjinP.R. China
  2. 2.College of Life ScienceNankai UniversityTianjinP. R. China
  3. 3.Department of Human Physiology and Center for NeuroscienceFlinders UniversityAdelaideAustralia

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