Immortalized human dorsal root ganglion cells differentiate into neurons with nociceptive properties.
A renewable source of human sensory neurons would greatly facilitate basic research and drug development. We had established previously conditionally immortalized human CNS cell lines that can differentiate into functional neurons (). We report here the development of an immortalized human dorsal root ganglion (DRG) clonal cell line, HD10.6, with a tetracycline-regulatable v-myc oncogene. In the proliferative condition, HD10.6 cells have a doubling time of 1.2 d and exhibit a neuronal precursor morphology. After differentiation of clone HD10.6 for 7 d in the presence of tetracycline, v-myc expression was suppressed, and >50% of the cells exhibited typical neuronal morphology, stained positively for neuronal cytoskeletal markers, and fired action potentials in response to current injection. Furthermore, this cell line was fate-restricted to a neuronal phenotype; even in culture conditions that promote Schwann cell or smooth muscle differentiation of neural crest stem cells, HD10.6 differentiated exclusively into neurons. Moreover, differentiated HD10.6 cells expressed sensory neuron-associated transcription factors and exhibited capsaicin sensitivity. Taken together, these data indicate that we have established an immortalized human DRG cell line that can differentiate into sensory neurons with nociceptive properties. The cell line HD10.6 represents the first example of a human sensory neuronal line and will be valuable for basic research, as well as for the discovery of novel drug targets and clinical candidates.
Raymon
H K
HK
Signal Pharmaceuticals Incorporated, San Diego, California 92121, USA.
Thode
S
S
Zhou
J
J
Friedman
G C
GC
Pardinas
J R
JR
Barrere
C
C
Johnson
R M
RM
Sah
D W
DW
D000200:Action Potentials; D002211:Capsaicin / Q000494:pharmacology; D002412:Cations / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D048429:Cell Size; D002999:Clone Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology* / Q000196:embryology; D016259:Genes, myc / Q000235:genetics; D006801:Humans; D015640:Ion Channel Gating; D008024:Ligands; D009475:Neurons, Afferent / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D009619:Nociceptors / Q000502:physiology*; D010146:Pain; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology; D014157:Transcription Factors / Q000032:analysis
D002412:Cations; D008024:Ligands; D014157:Transcription Factors; D013752:Tetracycline; D002211:Capsaicin