Mouse DRG Cell Line with Properties of Nociceptors

• Published in: PloS one Vol. 10; no. 6; p. e0128670
• Main Authors: Doran, Ciara, Chetrit, Jonathan, Holley, Matthew C, Grundy, David, Nassar, Mohammed A
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 08-06-2015; Public Library of Science (PLoS)
• Subjects: Animals; Antigens; Axonogenesis; Biomarkers - metabolism; Bradykinin; Calcium; Calcium channels (voltage-gated); Capsaicin; Capsaicin receptors; Cell culture; Cell Differentiation - drug effects; Cell Line, Transformed; Cell Lineage - drug effects; Cell lines; Cell Proliferation - drug effects; Cell Shape - drug effects; Clone Cells; Cloning; Drug delivery; Drug discovery; Ganglia, Spinal - cytology; Ganglia, Spinal - drug effects; Ganglia, Spinal - metabolism; Gene Expression Regulation - drug effects; Green Fluorescent Proteins - metabolism; Histamine; Immunocytochemistry; Immunohistochemistry; Inflammation; Inflammation Mediators - metabolism; Interferon; Markers; Mice; Microfilament Proteins - metabolism; Nervous system; Neurons; Neurosciences; Nociceptors; Nociceptors - metabolism; Norepinephrine; Pain; Pain perception; Polymerase chain reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Science; Sensory Receptor Cells - cytology; Sensory Receptor Cells - drug effects; Sensory Receptor Cells - metabolism; Sodium; Sodium channels (voltage-gated); Sodium Channels - metabolism; Stem cells; Transfection; Veratridine; Veratridine - pharmacology; United Kingdom > UK
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0128670

In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry, RT-PCR and calcium microfluorimetry, we demonstrate that the cell line MED17.11 expresses markers of cells committed to the sensory neuron lineage. Within a few hours under differentiating conditions, MED17.11 cells extend processes and following seven days of differentiation, express markers of more mature DRG neurons, such as NaV1.7 and Piezo2. However, at least at this time-point, the nociceptive marker NaV1.8 is not expressed, but the cells respond to compounds known to excite nociceptors, including the TRPV1 agonist capsaicin, the purinergic receptor agonist ATP and the voltage gated sodium channel agonist, veratridine. Robust calcium transients are observed in the presence of the inflammatory mediators bradykinin, histamine and norepinephrine. MED17.11 cells have the potential to replace or reduce the use of primary DRG culture in sensory, pain and developmental research by providing a simple model to study acute nociception, neurite outgrowth and the developmental specification of DRG neurons.