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cat no | ioEA1024 Early Access
ioSensory Neurons™
Highly pure human iPSC-derived sensory neurons with a defined nociceptor identity
ioSensory Neurons are a highly pure, easy to use and functional in vitro nociceptor model, enabling reliable chronic nociceptive pain research and therapeutics development for peripheral neuropathies.
ioSensory Neurons are precision reprogrammed using opti-ox™ technology, meaning consistency is built-in. Within 7 days post-revival, ioSensory Neurons form a highly pure (>99%) sensory neuronal population with a defined nociceptor identity, characterised by the expression of the pan-sensory neuron markers, PRPH, BRN3A, ISL1 and TUBB3, and key markers of nociceptors, NTRK1 and TRPV1.
These cells display spontaneous activity and show a functional nociceptor phenotype, as demonstrated by responsiveness to selective agonists for TRPV1, TRPM3, and TRPM8.
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Benchtop benefits
Highly Pure
>99% pure sensory neurons with a defined nociceptor identity by day 7 post-revival, as confirmed by single cell RNA sequencing.
Functional
ioSensory Neurons display spontaneous activity and a functional nociceptor phenotype, as shown by calcium mobilisation in response to specific TRP agonists.
Easy to use
Cryopreserved cells arrive, programmed to mature rapidly upon revival. Simple two-step protocol using one medium – no mitomycin C treatment is necessary.
Ready within days
opti-ox precision reprogrammed ioSensory Neurons rapidly form a homogenous neuronal population
Time-lapse video capturing the rapid and homogeneous neuronal phenotype acquisition upon thawing of cryopreserved ioSensory Neurons. 14 day time course.
Highly characterised and defined
ioSensory Neurons express pan-sensory neuron-specific markers
Immunofluorescent staining of ioSensory Neurons at day 14 post-revival. The upper panel shows that ioSensory Neurons are positive for BRN3A (red), the pan-neuronal marker MAP2 (green), and the DAPI counterstain (blue), and demonstrates that all MAP2 positive neurons have a sensory neuronal identity. The lower panel shows that ioSensory Neurons are positive for ISL1 (magenta), PRPH (red), the pan-neuronal marker TUBB3 (green), and the DAPI counterstain (blue).
ioSensory Neurons show visible neuronal networks by day 7
Upon reprogramming, rapid morphological changes are observed in the cells, with neurons identified by day 4 post-revival. Visible neuronal networks are observed by day 7 post-thaw. Images show day 1 to 14 post-thawing; 10X magnification; scale bar: 200 μm.
Single cell RNA-sequencing shows ioSensory Neurons form a pure population (>99%) of sensory neurons
Single cell RNA-sequencing analysis was performed with ioSensory Neurons at four specific timepoints (days 7, 10, 14 and 17). By day 7, the population has a distinct expression profile indicating a pure population (>99%) of post-mitotic sensory neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single cell RNA-sequencing shows ioSensory Neurons express key pan-sensory neuronal markers
Single cell RNA-sequencing analysis was performed with ioSensory Neurons at four specific timepoints (days 7, 10, 14 and 17). By day 7, the expression of pan-sensory neuron marker genes, ISL1, ISL2, PRPH, BRN3A, together with the pan-neuronal markers TUBB3 and MAP2, could be detected in post-mitotic sensory neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single cell RNA-sequencing shows ioSensory Neurons express key nociceptor markers
Single cell RNA-sequencing analysis was performed with ioSensory Neurons at four specific timepoints (days 7, 10, 14 and 17). By day 7, the expression of key nociceptor marker genes (NTRK1 and TRPV1) could be detected in post-mitotic sensory neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Rapid gain of functional activity
ioSensory Neurons display spontaneous activity that matures over time
Multi electrode array (MEA) recordings of ioSensory Neurons at days 6 and 17. The activity maps show firing rate (A), spike amplitude (B) and % of active electrodes (C). Results demonstrate a time-dependent increase of spontaneous activity during neuronal maturation from day 6 to day 17. Analysis was performed on a Maxwell Biosystem's MaxTwo multi-well system. Note, this data is from cells in continuous culture and not cryopreserved cells.
ioSensory Neurons display a functional nociceptor phenotype
Calcium mobilisation imaging upon stimulation of ioSensory Neurons with pharmacological agonists targeting thermosensitive TRP channels such as TRPV1 (capsaicin), TRPM3 (CIM0216) and TRPM8 (WS-12). Active traces represent the increase in intracellular calcium mobilisation of individual cells upon exposure to noxious stimuli but not to vehicle at day 17. This indicates that cells display a functional nociceptor phenotype within 17 days. Note, this data is from cells in continuous culture and not cryopreserved cells.
Calcium imaging of ioSensory Neurons demonstrates activity following treatment with a TRPM3 agonist
Representative video of ioSensory Neurons displaying calcium mobilisation in response to treatment with the TRPM3 agonist, CIM0216. Note, this data is from cells in continuous culture and not cryopreserved cells.
Cells arrive ready to plate
ioSensory Neurons are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: Induction, which is carried out at bit.bio (Phase 0), Stabilisation for 7 days (Phase 1), and Maintenance (Phase 2) during which the ioSensory Neurons mature. Phases 1 and 2 after revival of cells are carried out by the customer.
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