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Nat Protoc. 2016 Oct;11(10):1877-88. doi: 10.1038/nprot.2016.111. Epub 2016 Sep 8.

Surgical extraction of human dorsal root ganglia from organ donors and preparation of primary sensory neuron cultures.

Author information

1
Washington University Pain Center and Department of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri, USA.
2
Medical Scientist Training Program, Washington University in St. Louis, St. Louis, Missouri, USA.
3
Neurosciences Program, Washington University in St. Louis, St. Louis, Missouri, USA.
4
Pain Research Center, Department of Anesthesiology, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA.
5
Developmental, Regenerative, and Stem Cell Biology Program, Washington University in St. Louis, St. Louis, Missouri, USA.
6
Department of Neuroscience, Washington University School of Medicine, St. Louis, Missouri, USA.

Abstract

Primary cultures of rodent sensory neurons are widely used to investigate the cellular and molecular mechanisms involved in pain, itch, nerve injury and regeneration. However, translation of these preclinical findings may be greatly improved by direct validation in human tissues. We have developed an approach to extract and culture human sensory neurons in collaboration with a local organ procurement organization (OPO). Here we describe the surgical procedure for extraction of human dorsal root ganglia (hDRG) and the necessary modifications to existing culture techniques to prepare viable adult human sensory neurons for functional studies. Dissociated sensory neurons can be maintained in culture for >10 d, and they are amenable to electrophysiological recording, calcium imaging and viral gene transfer. The entire process of extraction and culturing can be completed in <7 h, and it can be performed by trained graduate students. This approach can be applied at any institution with access to organ donors consenting to tissue donation for research, and is an invaluable resource for improving translational research.

PMID:
27606776
PMCID:
PMC5082842
DOI:
10.1038/nprot.2016.111
[Indexed for MEDLINE]
Free PMC Article

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