Immortalization of Nociceptive Dorsal Root Ganglion Sensory Neurons
This invention involves a novel method to immortalize nociceptive dorsal root ganglion (DRG) neurons. DRG neurons are terminally differentiated cells that extend long axons to their target tissues. More than half of all DRG neurons are unmyelinated axons that extend to the skin and have nociceptive properties. In many studies on peripheral neuropathies and neuropathic pain, rat or mouse primary DRG neurons are used. However, obtaining these cells in sufficient quantities to perform high-throughput drug screening is nearly impossible. In order to immortalize rat DRG neurons, yet keep them in a state where they can stop dividing and differentiate into neurons with axons, we used a two-step transformation process. After double transformation, we are able to grow the cells through multiple doublings without loss of differentiation potential. These immortalized cells can be grown in large quantities, differentiated into nociceptive sensory neurons and can be used in high-throughput drug screening for neuropathic pain and peripheral neuropathy research. Background Most researchers, including large pharmaceutical or biotechnology companies working on neuropathic pain and peripheral neuropathies use dorsal root ganglion neurons as a model cell type to examine the effects of drugs on the biology of pain-sensitive (nociceptive) neurons. However, cell numbers that can be obtained are very limited with primary DRG neurons. It is almost impossible to do high-throughput drug screening with the currently available cells. An alternative is to use neuronal cell lines derived from neuroblastomas or other cancers but in this instance the issue is relevance to neuropathic pain research. These cells are often heterogeneous and do not respond to drugs in consistent manner. The nociceptive DRG neuronal cell lines that we have developed are clonal lines; therefore all of the cells are similar to each other and in biological assays behave in a predictable manner. These cells can be grown in large quantities, differentiated into nociceptive neurons that express proper markers and ion channels necessary for nociception and generation of action potentials, and used in high-throughput drug screening. Description (Set) Proposed Use (Set) This technology enables one to immortalize nociceptive dorsal root ganglion neurons to grow large quantities for use in high throughput drug screening for neuropathic pain and peripheral neuropathy research. The transfection and selection methods developed as part of this invention can also be used to generate other neuronal populations, including neurons from human tissues such as brain, spinal cord, dorsal root sensory ganglia and autonomic ganglia.Inventor(s): Hoke, Ahmet
Type of Offer: Licensing
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