Elsevier

Neuroscience

Volume 91, Issue 2, June 1999, Pages 519-525
Neuroscience

Neural transplantation of human neuroteratocarcinoma (hNT) neurons into ischemic rats. A quantitative dose–response analysis of cell survival and behavioral recovery

https://doi.org/10.1016/S0306-4522(98)00610-1Get rights and content

Abstract

Transplantation of fetal neuronal tissue has been used successfully to ameliorate symptoms of neurodegenerative disease in animals and humans. This technique has recently been extended as an experimental treatment for ischemic brain damage. However, due to ethical issues with the use of fetal cells for the treatment of any human disease, there has been a concerted effort to find alternative graft sources for neural transplantation. The human neuroteratocarcinoma neuron cell is derived from an embryonal teratocarcinoma cell line that can be differentiated into post-mitotic neurons. Neural transplantation of human neuroteratocarcinoma neurons has recently been shown to produce behavioral amelioration of symptoms in rats with ischemia-induced injury. The present study was undertaken to: (i) determine the minimum effective number of transplanted human neuroteratocarcinoma neurons required for amelioration of ischemia-induced behavioral dysfunction; and (ii) quantify the survival of human neuroteratocarcinoma neurons in vivo. Transplants of 0, 5, 10, 20, 40, 80 or 160×103 human neuroteratocarcinoma neurons were made into rats that sustained ischemic damage. Animals that received 40, 80 or 160×103 human neuroteratocarcinoma neurons demonstrated a dose-dependent improvement in performance of both the passive avoidance and elevated body swing tests. At the conclusion of behavioral testing, human neuroteratocarcinoma neurons were identified in paraffin sections with human neural cell adhesion molecule MOC-1 and human neurofilament antibodies. Transplants of 80 or 160×103 human neuroteratocarcinoma neurons demonstrated a 12–15% survival of human neuroteratocarcinoma neurons in the graft, while transplants of 40×103 human neuroteratocarcinoma neurons demonstrated a 5% survival.

Transplantation of human neuroteratocarcinoma neurons ameliorated behavioral deficits produced by ischemic damage. The human neuroteratocarcinoma neuron, additionally, showed greater survival than that reported for fetal cells when transplanted into the brain. Therefore, this readily available cell may prove to be an excellent candidate for the treatment of ischemic damage in human patients.

Section snippets

Experimental procedures

Animals for these studies were 85 male Sprague–Dawley (Zivic–Miller) rats. These animals received unilateral ischemic lesions to the striatum at eight weeks-of-age by subjecting them to unilateral occlusion of the middle cerebral artery for 1 h.5., 8., 21., 28. One month following lesion, the animals were behaviorally tested using a step-down passive avoidance and motor asymmetry test. Those animals that demonstrated behavioral deficits as a result of the ischemic lesion were randomly assigned

Behavioral results

Behavioral data from both the passive avoidance test and EBST demonstrated that learning and memory performance and motor behavior were impaired by the ischemic insult.

Behavioral recovery

Ischemia-induced dysfunction in passive avoidance learning and memory, and in asymmetrical motor behavior, were significantly improved in animals that received 40×103 hNT neurons or more. The behavioral recovery was seen as early as one month post-transplantation, and persisted throughout the entire three-month test period. The results replicated previous observations demonstrating that transplantation of 40×103 hNT neurons into the striatum of ischemically-damaged rats ameliorated behavioral

Conclusions

The readily available hNT neuron shows greater survival than fetal cells and promotes behavioral recovery in rats with ischemic damage. The hNT neuron, therefore, may prove to be an excellent candidate for the treatment of ischemic damage in human patients.

Acknowledgements

The authors would like to thank Stephan Labaste and Stacy Jones Sinibaldi for their expert technical assistance. This study was supported, in part, by Layton BioScience.

References (40)

  • M. Schinstine et al.

    Potential effect of cytokines on transgene expression in primary fibroblasts implanted into the rat brain

    Molec. Brain Res.

    (1997)
  • J.Q. Trojanowski et al.

    Neurons derived from a teratocarcinoma cell line establish molecular and structural polarity following transplantation into the rodent brain

    Expl Neurol.

    (1993)
  • M.J. West

    New stereological methods for counting neurons

    Neurobiol. Aging

    (1993)
  • P.W. Andrews

    Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro

    Devl Biol.

    (1987)
  • K. Bankiwicz et al.

    Trophism, transplantation and animal models of Parkinson's disease

    Expl Neurol.

    (1993)
  • A. Björklund

    Better cells for brain repair

    Nature

    (1993)
  • C.V. Borlongan et al.

    Behavioral recovery from focal ischemia after intrastriatal transplants of human-embryonal carcinoma-derived neurons (hNT cells)

    Int. neural Transplant. Meeting Abstr.

    (1997)
  • C.V. Borlongan et al.

    Elevated body swing test: a new behavioral parameter for rats with 6-hydroxydopamine-induced hemiparkinsonism

    J. Neurosci.

    (1995)
  • Borlongan C. V., Poulos S. G., Othberg A. and Sanberg P. R. Viability and survival of hNT neurons determine degree of...
  • I. Date et al.

    Cografting with polymer-encapsulated human nerve growth factor-secreting cells and chromaffin cell survival and behavioral recovery in hemiparkinsonian rats

    J. Neurosurg.

    (1996)
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    Present address: National Institute on Drug Abuse, Division of Intramural Research, 5500 Nathan Shock Drive, Baltimore, MD 21224, U.S.A.

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