The Journal of Pain
Volume 8, Issue 1 , Pages 33-50 , January 2007

Subarachnoid Transplant of a Human Neuronal Cell Line Attenuates Chronic Allodynia and Hyperalgesia After Excitotoxic Spinal Cord Injury in the Rat

  • Mary J. Eaton

      Affiliations

    • VA RR&D Center of Excellence in Functional Recovery in Chronic Spinal Cord Injury, VAMC, Miami, Florida.
    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • Department of Neurosurgery, Miller School of Medicine, University of Miami, Miami, Florida.
    • Corresponding Author InformationAddress reprint requests to Mary J. Eaton, PhD, Associate Professor of Neurological Surgery, The Miami Project to Cure Paralysis, Miller School of Medicine at the University of Miami, 1095 NW 14th Terrace (R-48), Miami, FL 33136.
    • ,
  • Stacey Quintero Wolfe

      Affiliations

    • Department of Neurosurgery, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Miguel Martinez

      Affiliations

    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Massiel Hernandez

      Affiliations

    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Cassandra Furst

      Affiliations

    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Jian Huang

      Affiliations

    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Beata R. Frydel

      Affiliations

    • The Miami Project to Cure Paralysis, Miller School of Medicine, University of Miami, Miami, Florida.
    • ,
  • Orlando Gómez-Marín

      Affiliations

    • VA RR&D Center of Excellence in Functional Recovery in Chronic Spinal Cord Injury, VAMC, Miami, Florida.
    • Department of Epidemiology & Public Health, Miller School of Medicine, University of Miami, Miami, Florida.

Received 27 February 2006 ,Revised 11 May 2006 ,Accepted 20 May 2006.

References 

  1. Andrews PW. The characteristics of cell lines derived from human germ cell tumors. In:  Damjanov I,  Knowles BB,  Solter D editor. The Human Teratomas: Experimental and Clinical Biology. Clifton, NJ: Humana; 1983;p. 285–311
  2. Andrews PW. Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol. 1984;103:285–293
  3. Andrews PW, Damjanov I, Simon D, Banting GS, Carlin C, Dracopoli NC, et al. Pluripotent embryonal carcinoma clones derived from human teratocarcinoma cell line Tera-2. Lab Invest. 1984;50:147–162
  4. Aran S, Hammond DL. Antagonism of baclofen-induced antinociception by intrathecal administration of phaclofen or 2-hydroxy-saclofen, but not delta-aminovaleric acid in the rat. J Pharm Exp Ther. 1991;257:360–368
  5. Bain G, Gottlieb DI. Neural cells derived by in vitro differentiation of P19 and embryonic stem cells. Perspectives Dev Neurobiol. 1998;5:175–178
  6. Balazy TE. Clinical management of chronic pain in spinal cord injury. Clin J Pain. 1992;8:102–110
  7. Berg-Johnsen J, Roste G, Solgaard T, Lundar T. Continuous intrathecal infusion of baclofen: A new therapeutic method for spasticity. Tidsskr Nor Laegeforen. 1998;118:3256–3260
  8. Bertman LJ, Advokat C. Comparison of the antinociceptive and antispastic action of (-)-baclofen after systemic and intrathecal administration in intact, acute and chronic spinal rats. Brain Res. 1995;684:8–18
  9. Brewer KL, Yezierski RP. Effects of adrenal medullary transplants on pain-related behaviors following excitotoxic spinal cord injury. Brain Res. 1998;798:83–92
  10. Brown D, Stanfield B. The use of bromodeoxyuridine-immunohistochemistry to identify transplanted fetal brain tissue. J Neural Transplant. 1989;1:135–139
  11. Canavero S, Bonicalzi V. The neurochemistry of central pain: Evidence from clinical studies, hypothesis and therapeutic implications. Pain. 1998;74:109–114
  12. Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Meth. 1994;53:55–63
  13. Cheung WM, Fu WY, Hui WS, Ip NY. Production of human CNS neurons from embryonal carcinoma cells using a cell aggregation method. BioTechniques. 1999;26:946–954
  14. Christensen MD, Pickelman J, Ziainia M, Hulsebosch CE. Temporal development of mechanical allodynia and thermal hyperalgesia in chronic central pain following spinal cord injury. Am Pain Soc Abs. 1995;95:811
  15. Daadi MM, Saporta S, Willing AE, Zigova T, McGrogan MP, Sanberg PR. In vitro induction and in vivo expression of bcl-2 in the hNT neurons. Brain Res Bull. 2001;56:147–152
  16. Dickenson AH, Chapman V, Green GM. The pharmacology of excitatory and inhibitory amino acid-mediated events in the transmission and modulation of pain in the spinal cord. Gen Pharm. 1997;28:633–638
  17. Dmitrovsky E, Moy D, Miller WH, Li A, Masui H. Retinoic acid causes a decline in TGF-alpha expression, cloning efficiency, and tumorigenicity in a human embryonal cancer cell line. Oncogene Res. 1990;5:233–239
  18. Donzanti BA, Yamamoto BK. An improved and rapid HPLC-EC method for the isocratic separation of amino acids neurotransmitters from brain tissue and microdialysis perfusates. Life Sci. 1988;43:913–922
  19. Dumoulin A, Rostaing P, Bedet C, Levi S, Isabert MF, Henry JP, et al. Presence of the vesicular inhibitory amino acid transporter in GABAergic and glycinergic synaptic terminal boutons. J Cell Sci. 1999;112:811–823
  20. Eaton MJ. Cell therapy for neuropathic pain in spinal cord injuries. Expert Opin Biol Ther. 2004;4:1861–1869
  21. Eaton MJ. Development of human cell therapy for functional recovery following SCI. J Spinal Cord Med. 2004;27:155
  22. Eaton MJ, Plunkett JA, Martinez MA, Karmally S, Montanez K, Rabade J. Changes in GAD and GABA immunoreactivity in the spinal dorsal horn after peripheral nerve injury and promotion of recovery by lumbar transplant of immortalized serotonergic neurons. J Chem Neuroanat. 1998;16:57–72
  23. Eaton MJ, Plunkett JA, Martinez MA, Lopez T, Karmally S, Cejas P, et al. Transplants of neuronal cells bio-engineered to synthesize GABA alleviate chronic neuropathic pain. Cell Transplant. 1999;8:87–101
  24. Eaton MJ, Sagen J. Cell therapy for models of pain and traumatic brain injury. In:  Sanberg PR,  Davis CD editor. Cell Therapy for Brain Repair. Totowa, NJ: The Humana Press, Inc; 2006;p. 199–239
  25. Follet KA, Naumann CP. A prospective study of catheter-related complications of intrathecal drug delivery systems. J Pain Symptom Manage. 2000;19:209–215
  26. Fromm GH. Baclofen as an adjuvant analgesic. J Pain Symptom Manage. 1994;9:500–509
  27. Globus MYT, Wester P, Busto R, Dietrich WD. Ischemia-induced extracellular release of serotonin plays a role in CA1 neuronal cell death. Stroke. 1992;23:1595–1601
  28. Gortz P, Fleischer W, Rosenbaum C, Otto FR, Siebler M. Neuronal network properties of human teratocarcinoma cell line-derived neurons. Brain Res. 2004;1018:18–25
  29. Gottlieb DI, Huettner JE. An in vitro pathway from embryonic stem cells to neurons and glia. Cells Tissues Organs. 1999;165:165–172
  30. Guillemain I, Alonoso G, Patey G, Privat A, Chaudieu I. Human NT2 neurons express a large variety of neurotransmission phenotypes in vitro. J Comp Neurol. 2000;422:380–395
  31. Hammond DL, Washington JD. Antagonism of L-baclofen-induced antinociception by CGP 35348 in the spinal cord of the rat. Eur J Pharmacol. 1993;234:255–262
  32. Hao JX, Xu XJ, Yu YX, Seiger A, Wiesenfeld-Hallin Z. Baclofen reverses the hypersensitivity of dorsal horn wide dynamic range neurons to mechanical stimulation after transient spinal cord ischemia: Implications for a tonic GABAergic inhibitory control of myelinated fiber input. J Neurophysiol. 1992;68:392–396
  33. Hao J-X, Xu X-J, Aldskogius H, Seiger A, Wiesenfeld-Hallin Z. Chronic pain related syndrome in rats after ischemic spinal cord lesion: A possible animal model for pain in patients with spinal cord injury. Pain. 1992;48:279–290
  34. Hargreaves K, Dubner R, Brown F, Flores C, Joris J. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain. 1988;32:77–88
  35. Heilporn A. Two therapeutic experiments on stubborn pain in spinal cord lesions: Coupling melitraceflupenthixol and transcutaneous nerve stimulation. Paraplegia. 1977;15:368–372
  36. Henry J. Pharmacological studies on the prolonged depressant effects of baclofen on lumbar dorsal horn units in the cat. Neuropharmacology. 1982;21:1085–1093
  37. Herman RM, D’Luzansky SC, Ippolito R. Intrathecal baclofen suppresses central pain in patients with spinal lesions: A pilot study. Clin J Pain. 1992;8:338–345
  38. Hulsebosch CE, Christensen MD, Peng YB, Willis WD. Central sensitization of wide dynamic range neurons in chronic central pain after spinal cord hemisection. Am Pain Soc Abs. 1995;95:812
  39. Ibuki T, Hama AT, Wang X-T, Pappas GD, Sagen J. Loss of GABA immunoreactivity in the spinal dorsal horn of rats with peripheral nerve injury and promotion of recovery by adrenal medullary grafts. Neuroscience. 1997;76:845–858
  40. Jain M, Armstrong RJ, Tyers P, Barker RA, Rosser AE. GABAergic immunoreactivity is predominant in neurons derived from expanded human neural precursors cells in vitro. Exp Neurol. 2003;182:113–123
  41. Johnston GAR, Hailstone M, Freeman C. Baclofen: Stereoselective inhibition of excitatory amino acid release. J Pharm Pharmacol. 1980;32:230–231
  42. Kingery WS, Fields RD, Kocsis JD. Diminished dorsal root GABA sensitivity following chronic peripheral nerve injury. Exp Neurol. 1988;100:478–490
  43. Kjaer M, Nielsen H. The analgesic effect of the GABA-agonist THIP in patients with chronic pain of malignant origin. Br J Clin Pharmacol. 1983;16:477–485
  44. Kondziolka D, Wechsler L, Goldstein S, Meltzer C, Thulborn KR, Gebel J, et al. Transplantation of cultured human neuronal cells for patients with stroke. Neurology. 2000;55:565–569
  45. Lee VM, Andrews PW. Differentiation of NTERA-2 clonal human embryonal carcinoma cells into neurons involves the induction of all three neurofilament proteins. J Neurosci. 1986;6:514–521
  46. Liu J, Wolfe D, Hao S, Huang S, Glorioso JC, Mata M, et al. Peripherally delivered glutamic acid decarboxylase gene therapy for spinal cord injury pain. Mol Ther. 2004;10:57–66
  47. Liu S, Dancausse HA, Yezierski RP. Non-NMDA receptor antagonists protect against neurotoxic spinal cord injury in the rat. Soc Neurosci Abstr. 1994;20:1534
  48. Liu S, Ruenes GL, Yezierski RP. NMDA and non-NMDA receptor antagonists protect against excitotoxic injury in the rat spinal cord. Brain Res. 1997;756:160–167
  49. Loubser PG, Akman NM. Effects of intrathecal baclofen on chronic spinal cord injury pain. J Pain Symptom Manage. 1996;12:241–247
  50. Maerz WJ, Baselga J, Reuter VE, Mellado B, Myers ML, Bosl GJ, et al. FGF4 dissociates anti-tumorigenic from differentiation signals of retinoic acid in human embryonal carcinomas. Oncogene. 1998;17:761–767
  51. Malcangio M, Bowery N. Spinal cord SP release and hyperalgesia in monoarthritic rats: Involvement of the GABA B receptor system. Br J Pharmacol. 1994;113:1561–1566
  52. Marsala M, Kakinohana O, Yaksh TL, Tomori Z, Marsala S, Cizkova D. Spinal implantation of hNT neurons and neuronal precursors: Graft survival and functional effects in rats with ischemic spastic paraplegia. Eur J Neurosci. 2004;20:2401–2414
  53. Megiorni F, Mora B, Indovina P, Mazzilli MC. Expression of neuronal markers during NTera2/cloneD1 differentiation by cell aggregation method. Neurosci Lett. 2005;373:105–109
  54. Meltzer CC, Kondziolka D, Villermagne VL, Wechsler L, Goldstein S, Thulborn KR, et al. Serial [18F] fluorodeoxyyglucose positron emission tomography after human neuronal implantation for stroke. Neurosurgery. 2001;49:586–592
  55. Milhorat TH, Kotzen RM, Mu HT, Capocelli AL, Milhorat RH. Dysesthetic pain in patients with syringomyelia. Neurosurgery. 1996;38:940–946
  56. Miyazono M, Nowell PC, Finan JL, Lee VM, Trojanowski JQ. Long-term integration and neuronal differentiation of human embryonal carcinoma cells (NTera-2) transplanted into the caudoputamen of nude mice. J Comp Neurol. 1996;376:603–613
  57. Morrow TJ, Paulson PE, Brewer KL, Yezierski RP, Casey KL. Chronic, selective forebrain responses to excitotoxic dorsal horn injury. Exp Neurol. 2000;161:220–226
  58. Mouton PR. Principles and Practices of Unbiased Stereology: An introduction for bioscientists. Baltimore, MD: Johns Hopkins Press; 2002;
  59. Neelands TR, Zhang J, Macdonald RL. GABA (A) receptors expressed in undifferentiated human teratocarcinoma NT2 cells differ from those expressed by differentiated NT2-N cells. J Neurosci. 1999;19:7057–7065
  60. Nelson PT, Kondziolka D, Wechsler L, Goldstein S, Gebel J, DeCesare S, et al. Clonal human (hNT) neuron grafts for stroke therapy: neuropathology in a patient 27 months after implantation. Am J Pathol. 2002;160:1201–1206
  61. Newman MB, Misiuta I, Willing AE, Zigova T, Karl RC, Borlongan CV, et al. Tumorgenicity issues of embryonic carcinoma-derived stem cells: Relevance to surgical trials using NT2 and hNT neural cells. Stem Cells Devel. 2005;14:29–43
  62. Nguyen L, Rigo JM, Rocher V, Belachew S, Malgrange B, Rogister B, et al. Neurotransmitters as early signals for central nervous system development. Cell Tiss Res. 2001;305:187–202
  63. Pleasure SJ, Lee VM. NTera 2 cells: A human cell line which displays characteristics expected of a human committed neuronal progenitor cell. J Neurosci Res. 1993;35:585–602
  64. Pleasure SJ, Page C, Lee VM. Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci. 1992;12:1802–1815
  65. Richardson RR, Meyer PR, Cerullo LJ. Transcutaneous electrical neurostimulation in musculoskeletal pain of acute spinal cord injuries. Spine. 1980;8:75–84
  66. Saporta S, Willing AE, Colina LO, Zigova T, Milliken M, Daadi MM, et al. In vitro and in vivo characterization of hNT neuron neurotransmitter phenotypes. Brain Res Bull. 2000;53:263–268
  67. Satoh J, Kuroda Y. Differential gene expression between human neurons and neuronal progenitor cells in culture: An analysis of arrayed cDNA clones in NTera2 human embryonal carcinoma cell line as a model system. J Neurosci Methods. 2000;94:155–164
  68. Schwartz ED, Yezierski RP, Pattany PM, Quencer RM, Weaver RG. Diffusion-weighted MR imaging in a rat model of syringomyelia after excitotoxic spinal cord injury. Am J Neuroradiol. 1999;20:1422–1428
  69. Shapiro S. Neurotransmission by neurons that use serotonin, noradrenaline, glutamate, glycine, and gamma-aminobutyric acid in the normal and injured spinal cord. Neurosurgery. 1997;40:168–176
  70. Shihabuddin LS, Hertz JA, Holets VR, Whittemore SR. The adult CNS retains the potential to direct region-specific differentiation of a transplanted neuronal precursor cell line. J Neurosci. 1995;15:6666–6678
  71. Slonimski M, Abram SE, Zuniga RE. Intrathecal baclofen in pain management. Reg Anesth Pain Med. 2004;29:269–276
  72. Smith DF. Stereoselectivity of spinal neurotransmission: Effects of baclofen enantiomer on tail-flick reflex in rats. J Neur Trans. 1984;60:63–67
  73. Stiller CO, Cui JG, O’Connor WT, Brodin E, Meyerson BA, Linderoth B. GABA-ergic mechanisms may be involved in the spinal cord stimulation induced reversion of allodynia: Animal studies with microdialysis. Am Pain Soc Abs. 1995;95:801
  74. Supplisson S, Roux MJ. Why glycine transporters have different stoichiometries. FEBS Letts. 2002;529:93–101
  75. Taira T, Kawamura H, Tanikawa T, Iseki H, Kawabatake H, Takakura K. A new approach to control central deafferentation pain: spinal intrathecal baclofen. Stereotact Funct Neurosurg. 1995;65:101–105
  76. Todd AJ, McKenzie J. GABA-immunoreactive neurons in the dorsal horn of the rat spinal cord. Neuroscience. 1989;31:799–806
  77. Trojanowski JQ, Kleppner SR, Hartley RS, Miyazono M, Fraser NW, Kesari S, et al. Transfectable and transplantable postmitotic human neurons: Potential “platform” for gene therapy of nervous system diseases. Exp Neurol. 1997;144:92–97
  78. Trojanowski JQ, Mantione JR, Lee JH, Seid DP, You T, Inge LJ, et al. Neurons derived from a human tetratocarcinoma cell line establish molecular and structural polarity following transplantation into the rodent brain. Exp Neurol. 1993;122:283–294
  79. Varju P, Katarova Z, Madarasz E, Szabo G. GABA signalling during development: New data and old questions. Cell Tiss Res. 2001;305:239–246
  80. White LA, Eaton MJ, Castro MC, Klose KJ, Globus MY, Shaw G, et al. Distinct regulatory pathways control neurofilament expression and neurotransmitter synthesis in immortalized serotonergic neurons. J Neurosci. 1994;14:6744–6753
  81. Widerstrom-Noga EG, Felipe-Cuervo E, Broton JG, Duncan RC, Yezierski RP. Perceived difficulty in dealing with consequences of spinal cord injury. Arch Phys Med Rehabil. 1999;80:580–586
  82. Xu X-J, Hao J-X, Aldoskogius H, Seiger A, Wiesenfeld-Hallin Z. Chronic pain-related syndrome in rats after ischemic spinal cord lesion: A possible animal model for pain in patients with spinal cord injury. Pain. 1992;48:279–290
  83. Yaksh TL. Behavioral and autonomic correlates of the tactile evoked allodynia produced by spinal glycine inhibition: Effects of modulatory receptor systems and excitatory amino acid antagonists. Pain. 1989;37:111–123
  84. Yang L, Jones NR, Stoodley Ma, Blumbergs PC, Brown CJ. Excitotoxic model of post-traumatic syringomyelia in the rat. Spine. 2001;26:1842–1849
  85. Yezierski RP. Pain following spinal cord injury: The clinical problem and experimental studies. Pain. 1996;68:185–194
  86. Yezierski RP, Liu S, Ruenes GL, Kajander KJ: Behavioral and pathological characteristics of a central pain model following spinal injury. Presented at VIIIth World Congress on Pain, August 17-22, 1996, Vancouver, British Columbia, Canada
  87. Yezierski RP, Liu S, Ruenes GL, Kajander KJ, Brewer KL. Excitotoxic spinal cord injury: Behavioral and morphological characteristics of a central pain model. Pain. 1998;75:141–155
  88. Yezierski RP, Santana M, Park SH, Madsen PW. Neuronal degeneration and spinal cavitation following intraspinal injections of quisqualic acid in the rat. J Neurotrauma. 1993;10:445–456
  89. Yong VW, Kim SU. A new double labelling immunofluorescence technique for the determination of proliferation of human astrocytes in culture. J Neurosci Meth. 1987;21:9–16
  90. Yoshioka A, Yudkoff M, Pleasure D. Expression of glutamic acid decarboxylase during human neuronal differentiation: Studies using the NTera-2 culture system. Brain Res. 1997;767:333–339
  91. Yu CG, Fairbanks CA, Wilcox GL, Yezierski RP. Effects of agmatine, interleukin-10, and cyclosporin on spontaneous pain behavior after excitotoxic spinal cord injury in rats. J Pain. 2003;4:129–140
  92. Zhang A-L, Hao J-X, Seiger A, Xu X-J, Wiesenfeld-Hallin Z, Grant G, et al. Decreased GABA immunoreactivity in spinal cord dorsal horn neurons after transient spinal cord ischemia in the rat. Brain Res. 1994;656:187–190
  93. Zhang S, Tang J, Yuan B, Jia H. Involvement of the frontal ventrolateral orbital cortex in descending inhibition of nociception mediated by the periaqueductal gray in rats. Neurosci Lett. 1997;224:142–146

 Supported by the Department of Veterans Affairs Rehabilitation Research and Development Merit Review Grant; the VA RR&D Center of Excellence in Functional Recovery in Chronic Spinal Cord Injury, Miami VAMC; as well as grants from the American Syringomyelia Alliance Project; a grant from the Column of Hope, Chiari and Syringomyelia Research Foundation; and continuing support from the Miami Project to Cure Paralysis, Miller School of Medicine, Miami, Florida.

PII: S1526-5900(06)00844-3

doi: 10.1016/j.jpain.2006.05.013

The Journal of Pain
Volume 8, Issue 1 , Pages 33-50 , January 2007

Article Tools

* Image Usage & Resolution