title: Mouse DRG Cell Line with Properties of Nociceptors.
abstract: 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.
authors.affiliation: Department of Biomedical Science, University of Sheffield, United Kingdom.
authors.forename: Ciara
authors.initials: C
authors.lastname: Doran
authors.affiliation: Department of Biomedical Science, University of Sheffield, United Kingdom.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Chetrit
authors.affiliation: Department of Biomedical Science, University of Sheffield, United Kingdom.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Holley
authors.affiliation: Department of Biomedical Science, University of Sheffield, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Grundy
authors.affiliation: Department of Biomedical Science, University of Sheffield, United Kingdom.
authors.forename: Mohammed A
authors.initials: MA
authors.lastname: Nassar
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002461:Cell Line, Transformed; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002999:Clone Cells; D005727:Ganglia, Spinal / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D018836:Inflammation Mediators / Q000378:metabolism; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D009619:Nociceptors / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism; D014162:Transfection; D014701:Veratridine / Q000494:pharmacology
chemical_list: C113762:Avil protein, mouse; D015415:Biomarkers; D018836:Inflammation Mediators; D008840:Microfilament Proteins; D012333:RNA, Messenger; D015222:Sodium Channels; D049452:Green Fluorescent Proteins; D014701:Veratridine