Harvested human neurons engineered as live nervous tissue constructs: implications for transplantation

Laboratory investigation

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  • 1 Department of Neurosurgery, University of Rochester Medical Center, Rochester, New York;
  • 2 Department of Neurosurgery, University of Pennsylvania, Hospital of the University of Pennsylvania and
  • 3 Department of Pediatrics, University of Pennsylvania, The Children's Hospital of Philadelphia, Pennsylvania
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Object

Although neuron transplantation to repair the nervous system has shown promise in animal models, there are few practical sources of viable neurons for clinical application and insufficient approaches to bridge extensive nerve damage in patients. Therefore, the authors sought a clinically relevant source of neurons that could be engineered into transplantable nervous tissue constructs. The authors chose to evaluate human dorsal root ganglion (DRG) neurons due to their robustness in culture.

Methods

Cervical DRGs were harvested from 16 live patients following elective ganglionectomies, and thoracic DRGs were harvested from 4 organ donor patients. Following harvest, the DRGs were digested in a dispase–collagenase treatment to dissociate neurons for culture. In addition, dissociated human DRG neurons were placed in a specially designed axon expansion chamber that induces continuous mechanical tension on axon fascicles spanning 2 populations of neurons originally plated ∼ 100 μm apart.

Results

The adult human DRG neurons, positively identified by neuronal markers, survived at least 3 months in culture while maintaining the ability to generate action potentials. Stretch-growth of axon fascicles in the expansion chamber occurred at the rate of 1 mm/day to a length of 1 cm, creating the first engineered living human nervous tissue constructs.

Conclusions

These data demonstrate the promise of adult human DRG neurons as an alternative transplant material due to their availability, viability, and capacity to be engineered. Also, these data show the feasibility of harvesting DRGs from living patients as a source of neurons for autologous transplant as well as from organ donors to serve as an allograft source of neurons.

Abbreviations used in this paper: CGRP = calcitonin gene related peptide; DRG = dorsal root ganglion; PBS = phosphate-buffered saline.

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Contributor Notes

Address correspondence to: Douglas H. Smith, M.D., Department of Neurosurgery, 105 Hayden Hall, 3320 Smith Walk, University of Pennsylvania, Philadelphia, Pennsylvania 19104. email: smithdou@mail.med.upenn.edu.
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