Isolation and Long-term Survival of Adult Human Sensory Neurons In Vitro

Sosa, Iván J.; Reyes, Onix; Inserni, Jaime; Kuffler, Damién P.

doi:10.1097/00006123-199803000-00054

Cited by 13 publications

(15 citation statements)

References 14 publications

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“…Under optimized culturing conditions, we found that these neurons could survive in culture for at least 3 months, supporting previous observations of long-term survival. 11,14,23,24 Moreover, we have demonstrated that harvested DRG neurons can be engineered to create the first living human nervous tissue constructs using a newly identified process of axon growth. To form the constructs, axon tracts spanning 2 populations of DRG neuron bodies were stretch-grown from the initial length of ~ 100 μm and extended 10 mm in length, while maintaining normal morphological characteristics.…”

Section: Discussionmentioning

confidence: 99%

“…Kim and colleagues 11 maintained dissociated neurons from human superior cervical ganglia in vitro for > 2 months and demonstrated that action potentials could be recorded extracellularly from these neurons. Likewise, Sosa et al 23 were able to isolate DRG neurons from human organ donors and maintain them for 2.5 months and record action potentials after 8 weeks in culture. Finally, Baumann and colleagues 3 performed extensive electrophysiological examination of DRG neurons in relation to neuropathic pain with survival up to 90 days in culture.…”

Section: Discussionmentioning

confidence: 99%

“…4,6,10,19,25 In addition, several groups have successfully cultured DRG neurons from adult human organ donors, human fetuses, and recently from patients surgically treated with ganglionectomies to evaluate human DRG neuron physiology. 3,11,23,24 Dorsal root ganglia also appear to be an ideal source of neurons to engineer transplantable living nervous tissue constructs. We have recently shown that bundles of axons spanning 2 populations of rat DRG neuronal cell bodies readily tolerate extreme axon stretch-growth, creating living nerve tracts in vitro.…”

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confidence: 99%

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

Huang

Zager

Zhang

et al. 2008

JNS

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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. Stretchgrowth 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.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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

Huang

Zager

Zhang

et al. 2008

JNS

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“…Furthermore, a unique advantage of DRG neurons for clinical application is their availability. DRGs can be harvested from organ donors for allografts or from the patient themselves to create autografts (Maddox et al, 2004;Sosa et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Development of transplantable nervous tissue constructs comprised of stretch-grown axons

Pfister

Iwata

Taylor

et al. 2006

Journal of Neuroscience Methods

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“…Localized hypothermia (32°C) provides good neuroprotection during periods of compromised cerebral blood flow and oxygen delivery 49. Different studies show that the temperature that provides best neuroprotection varies considerably from mild (33°C–35°C)50 to moderate (30°C–32°C),51 severe (27°C–29°C),51 and extreme (20°C) 52…”

Section: Neuroprotectionmentioning

confidence: 99%

Maximizing neuroprotection: where do we stand?

Kuffler¹

2012

TCRM

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Brain and spinal cord traumas include blunt and penetrating trauma, disease, and required surgery. Such traumas trigger events such as inflammation, infiltration of inflammatory and other cells, oxidative stress, acidification, excitotoxicity, ischemia, and the loss of calcium homeostasis, all of which cause neurotoxicity and neuron death. To prevent trauma-induced neurological deficits and death, each of the many neurotoxic events that occur in parallel or sequentially must be minimized or prevented. Although neuroprotective techniques have been developed that block single neurotoxic events, most provide only limited neuroprotection and are only applied singly. However, because many neurotoxicity triggers arise from common events, an approach for invoking more effective neuroprotection is to apply multiple neuroprotective methods simultaneously before the many neurotoxic triggers and cascades are initiated and become irreversible. This paper first discusses some triggers of neurotoxicity and neuroprotective mechanisms that block them, including hypothermia, alkalinization, and the administration of adenosine. It then examines how the simultaneous application of these techniques provides significantly greater neuroprotection than is provided by any technique alone. The paper also stresses the importance of determining whether the neuroprotection provided by these techniques can be further enhanced by combining them with additional techniques, such as the systemic administration of glucocorticoids. Finally, the paper stresses the absolute critical importance of applying these techniques within the “golden hour” following trauma, before the many neurotoxic events and cascades are manifest and before the neurotoxic cascades become irreversible.

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Isolation and Long-term Survival of Adult Human Sensory Neurons In Vitro

Cited by 13 publications

References 14 publications

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

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

Development of transplantable nervous tissue constructs comprised of stretch-grown axons

Maximizing neuroprotection: where do we stand?

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