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An improved preservation method for human dorsal root ganglion neurons enables wider access to human molecular pain neuroscience.

Overview

Authors: Joseph B. Lesnak1, Mandee K. Schaub1, Kimberly Gomez2, Aida Calderon-Rivera2, Santiago Loya-Lopez2, Robert Stewart3, Sooyeon Jo3, Akie Fujita3, Tomás Osorno3, Hemanth Mydugolam1, Marisa Desai1, Keerthana Natarajan1, Morgan K. Schackmuth1, Marisol Mancilla Moreno1, Stephanie I. Shiers1, Anna Cervantes4, Geoffrey Funk4, Peter Horton4, Erin Vines4, Muhammad Saad Yousuf1, Katelyn E. Sadler1, Bruce P. Bean3, Rajesh Khanna2, Gregory Dussor1, Theodore J. Price1
ORCID iDs: Joseph B. Lesnak, Theodore J. Price
  1. Department of Neuroscience, Center for Advanced Pain Studies, The University of Texas at Dallas, Richardson, TX, USA
  2. Department of Pharmacology and Therapeutics, Center for Advanced Pain Therapeutics and Research (CAPToR), College of Medicine, University of Florida, Gainesville, FL, USA
  3. Department of Neurobiology, Harvard Medical School, Boston, MA, USA
  4. Southwest Transplant Alliance, Dallas, TX, USA
Institutions: The University of Texas at Dallas (United States); University of Florida Health (United States); University of Florida (United States); Harvard University (United States); Southwest Transplant Alliance (United States)
Journal: Cell reports methods, volume 6, issue 5, article 101412
Dates: received 14 September 2025; accepted 23 March 2026; published online 17 April 2026; in print May 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1016/j.crmeth.2026.101412 · PMID 41999764 · PMCID PMC13198098 · OpenAlex W7154736860
Open access: gold, a free copy (OpenAlex)
Status: code on request
Categories: optical imaging (calcium, voltage, 2-photon) (modality), human (organism), pain (population), cellular / molecular (subfield)
Methods: Statistics, Single-unit activity, calcium imaging
Keywords: Hibernate A, dorsal root ganglia, neurons, nociceptors, human tissue, electrophysiology, calcium imaging, tissue preservation
MeSH: Ganglia, Spinal*, Neurons*, Neurosciences*, Pain*, Tissue Preservation*, Cells, Cultured, Humans, Nociceptors (* major topic)
Topic: Pain Mechanisms and Treatments (Physiology, Medicine), according to OpenAlex
Funding: U.S. Department of Health & Human Services | NIH | National Institute of Neurological Disorders and Stroke (NINDS); National Institutes of Health (U19NS130608, R01NS111929, F32NS134563, R35-NS127216, RF1NS131165, K99NS134965); Department of Defense (HT9425-24-1-1007, CP230147P1)
Citations: cited by 3 papers (Europe PMC); 27 references in the paper
Research resources: Chicken anti-Peripherin RRID:AB_2284443, Goat anti-Chicken IgY Alexa Fluor 488 RRID:AB_2534096, CD45 antibody RRID:AB_2562106, TruStain FcX (human Fc block) RRID:AB_28118986, CD3 antibody RRID:AB_314065, CD11b antibody RRID:AB_830644, RRID:SCR_025458

Abstract

The use of human dorsal root ganglion (DRG) from organ donors opens the door for research into molecular biology and physiology of human nociceptors; however, there are barriers to working with this tissue including logistical difficulties and limited access. We present an approach using Hibernate A media to store whole DRGs or dissociated neurons prior to culturing and functional testing. Dissociation of DRGs following temporary storage (4–16 h) in Hibernate A media resulted in similar neuronal and immune cell yield as acutely dissociated DRGs. Neurons derived from DRGs stored in Hibernate A media prior to dissociation exhibited similar electrophysiological properties and capsaicin responses as acutely dissociated DRG neurons. Similarly, neurons from acutely dissociated DRGs stored in Hibernate A media (16–42 h) and shipped to geographically distant laboratories produced neuronal cultures displaying comparable electrophysiological properties as acutely cultured neurons. This approach overcomes insurmountable logistical burdens and increases access to freshly recovered human DRGs.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

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Data

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Data and code availability

• All data reported in this paper will be shared by the lead contact upon request. • No new original code was created in this paper. • Additional information required to reanalyze the data reported in this paper is available from the lead contact upon request.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

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Version 1, 29 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 25 authors, 8 keywords, 8 MeSH terms, 3 funders, 27 references, 7 RRIDs.

Cite

This paper

Lesnak, J. B., Schaub, M. K., Gomez, K., Calderon-Rivera, A., Loya-Lopez, S., Stewart, R., Jo, S., Fujita, A., Osorno, T., Mydugolam, H., Desai, M., Natarajan, K., Schackmuth, M. K., Moreno, M. M., Shiers, S. I., Cervantes, A., Funk, G., Horton, P., Vines, E., . . . Price, T. J. (2026). An improved preservation method for human dorsal root ganglion neurons enables wider access to human molecular pain neuroscience. Cell reports methods, 6(5), 101412. https://doi.org/10.1016/j.crmeth.2026.101412

BibTeX

@article{lesnak2026improved,
author = {Lesnak, Joseph B. and Schaub, Mandee K. and Gomez, Kimberly and Calderon-Rivera, Aida and Loya-Lopez, Santiago and Stewart, Robert and Jo, Sooyeon and Fujita, Akie and Osorno, Tomás and Mydugolam, Hemanth and Desai, Marisa and Natarajan, Keerthana and Schackmuth, Morgan K. and Moreno, Marisol Mancilla and Shiers, Stephanie I. and Cervantes, Anna and Funk, Geoffrey and Horton, Peter and Vines, Erin and Yousuf, Muhammad Saad and Sadler, Katelyn E. and Bean, Bruce P. and Khanna, Rajesh and Dussor, Gregory and Price, Theodore J.},
title = {{An improved preservation method for human dorsal root ganglion neurons enables wider access to human molecular pain neuroscience}},
journal = {Cell reports methods},
year = {2026},
month = apr,
volume = {6},
number = {5},
pages = {101412},
publisher = {Elsevier},
issn = {2667-2375},
doi = {10.1016/j.crmeth.2026.101412},
url = {https://doi.org/10.1016/j.crmeth.2026.101412},
pmid = {41999764},
pmcid = {PMC13198098}
}

RIS

TY - JOUR
AU - Lesnak, Joseph B.
AU - Schaub, Mandee K.
AU - Gomez, Kimberly
AU - Calderon-Rivera, Aida
AU - Loya-Lopez, Santiago
AU - Stewart, Robert
AU - Jo, Sooyeon
AU - Fujita, Akie
AU - Osorno, Tomás
AU - Mydugolam, Hemanth
AU - Desai, Marisa
AU - Natarajan, Keerthana
AU - Schackmuth, Morgan K.
AU - Moreno, Marisol Mancilla
AU - Shiers, Stephanie I.
AU - Cervantes, Anna
AU - Funk, Geoffrey
AU - Horton, Peter
AU - Vines, Erin
AU - Yousuf, Muhammad Saad
AU - Sadler, Katelyn E.
AU - Bean, Bruce P.
AU - Khanna, Rajesh
AU - Dussor, Gregory
AU - Price, Theodore J.
TI - An improved preservation method for human dorsal root ganglion neurons enables wider access to human molecular pain neuroscience
T2 - Cell reports methods
J2 - Cell Rep Methods
PY - 2026
DA - 2026/04/17
VL - 6
IS - 5
SP - 101412
SN - 2667-2375
PB - Elsevier
DO - 10.1016/j.crmeth.2026.101412
UR - https://doi.org/10.1016/j.crmeth.2026.101412
LA - en
ER -

CSL-JSON

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