cat no | io1024
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 deterministically programmed 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.
Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
per vial
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@bit.bio.
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.
opti-ox programmed 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.
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 deterministic programming, 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.
Single cell RNA-sequencing shows ioSensory Neurons form a pure population (>99%) of sensory neurons and express key pan-sensory neuronal markers
Single cell RNA-sequencing shows ioSensory Neurons express key nociceptor markers
Single cell RNA-sequencing analysis was performed with ioSensory Neurons at five specific timepoints: day 0 (iPSCs), 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 single cell RNA-sequencing. Note, this data is from cells in continuous culture, so minor variations may exist between this data and data from cryopreserved cells.
Bulk RNA sequencing analysis was performed on three technical replicates of three independent lots of ioSensory Neurons at different time points throughout the reprogramming protocol. Principal component analysis represents the variance in gene expression between the lots of ioSensory Neurons. This analysis shows high consistency between each lot of ioSensory Neurons at each given timepoint. Differential gene expression was found to be less than 1% between lots, at day 14 post-thaw. Pure populations of ioSensory Neurons with equivalent expression profiles can be generated consistently from every vial, allowing confidence in experimental reproducibility. Expression levels for specific genes of interest can be requested by contacting our team at technical@bit.bio.
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 enhanced TRPM3 and TRPM8 responses with bespoke media for these channels and increased culture length
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 two-phase process: Induction, which is carried out at bit.bio, 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.
Starting material
Human iPSC line
Karyotype
Normal (46, XY)
Seeding compatibility
6, 12, 24, 96 and 384 well plates
Shipping info
Dry ice
Donor
Caucasian adult male (skin fibroblast)
Vial size
Small: >2 x 10⁶ viable cells
Quality control
Sterility, protein expression (ICC) and gene expression (RT-qPCR)
Differentiation method
opti-ox deterministic cell programming
Recommended seeding density
60,000 cells/cm²
User storage
LN2 or -150°C
Format
Cryopreserved cells
Product use
ioCells are for research use only
Applications
Chronic pain research & drug development
MEA analysis
Calcium imaging
Transcriptome analysis
Neurotoxicology
Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | bit.bio
Innovation showcase talk at ISSCR
Marius Wernig MD, PhD | Stanford
Mark Kotter, MD, PhD | bit.bio
Download this infographic to find out how the approach used to generate human iPSC-derived cells influences purity, batch consistency and protocol speed.