ICC imaging of ioSensory Neurons - consistent, pure and easy to use sensory neurons for animal-free toxicology studies and chronic pain research

cat no | io1024

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.

Place your order

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

Benchtop benefits

pure_0

Highly Pure

>99% pure sensory neurons with a defined nociceptor identity by day 7 post-revival, as confirmed by single cell RNA sequencing.

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Functional

ioSensory Neurons display spontaneous activity and a functional nociceptor phenotype, as shown by calcium mobilisation in response to specific TRP agonists.

easy_0

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.

Technical data

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 

bitbio-ioSensory_Neurons-ICC_Comparison V2

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

bitbio-ioSensory_Neurons-Morphology_Timeline-correction

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.

Single cell RNA-sequencing shows ioSensory Neurons form a pure population (>99%) of sensory neurons and express key pan-sensory neuronal markers

FINAL scRNA Seq pan sensory marker figure_Morse
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 population has a distinct expression profile indicating a pure population (>99%) of post-mitotic sensory neurons, demonstrated through the expression of pan-sensory neuronal marker genes, ISL1, ISL2, PRPH, and BRN3A, together with the pan-neuronal markers TUBB3 and MAP2. 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.

Single cell RNA-sequencing shows ioSensory Neurons express key nociceptor markers

FINAL scRNA Seq nociceptor figure_Morse

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.

Lot-to-lot consistency

Whole transcriptome analysis shows high lot-to-lot consistency of ioSensory Neurons
FINAL Bulk RNA seq PCA plot_Morse

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.

Rapid gain of functional activity

ioSensory Neurons display spontaneous activity that matures over time
bitbio-ioSensory_Neurons-MEA_Data

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 traces_day 14_compressed
Calcium mobilisation imaging upon stimulation of ioSensory Neurons with pharmacological agonists targeting thermosensitive TRP channels such as TRPV1 (capsaicin), TRPM3 (CIM0216) and TRPM8 (WS12). Active traces represent the increase in intracellular calcium mobilisation of individual cells upon exposure to noxious stimuli but not to vehicle at day 14 post-revival. This indicates that cells display a functional nociceptor phenotype within 14 days post-thaw.
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.

ioSensory Neurons display enhanced TRPM3 and TRPM8 responses with bespoke media for these channels and increased culture length

TRPM3 and TRPM8 response with optimised media_compressed
Calcium mobilisation imaging, performed with ioSensory Neurons cultured using a bespoke media up to day 14, 17 or 21 post-thaw, shows that ioSensory Neurons display strong responses to pharmacological agonists targeting thermosensitive TRP channels, TRPM3 (CIM0216) and TRPM8 (WS12). The top and bottom left graphs show active traces which represent the increase in intracellular calcium mobilisation of individual cells, at day 21 post-thaw, upon exposure to noxious agonists but not to vehicle, indicating that cells display features of functional nociceptors. The bottom right graph shows the percentage of responding cells at day 14, 17, or 21 post-thaw – the bespoke media and increased culture length appears to greatly enhance the percentage of cells responding to the TRPM3 and TRPM8 agonists, to 90% and 67% at day 21 post-thaw, respectively. Please contact us at technical@bit.bio for further information on the bespoke media.

Easy culturing

Cells arrive ready to plate

bit.bio_ioSensory Neurons_timeline_horizontal

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.

Product information

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 cellular reprogramming

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

Product resources

ioSensory Neurons™ Brochure
ioSensory Neurons™
bit.bio
Download
ioSensory Neurons™ - User Manual User manual
ioSensory Neurons™ - User Manual
V2
2024
bit.bio
Download
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming Webinar
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming

Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | bit.bio

Watch now
Industrialising Cellular Reprogramming: Leveraging opti-ox™ Technology to Manufacture Human Cells with Unprecedented Consistency Talk
Industrialising Cellular Reprogramming: Leveraging opti-ox™ Technology to Manufacture Human Cells with Unprecedented Consistency

Innovation showcase talk at ISSCR

Marius Wernig MD, PhD | Stanford 

Mark Kotter, MD, PhD | bit.bio

Watch now

Differentiating iPSCs - which approach works best? 

Download this infographic to find out how the approach used to generate human iPSC-derived cells influences purity, batch consistency and protocol speed. 

bit.bio-opti-ox-infographic in hand-Aug 2022

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Related pages

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Resources Explore our latest scientific insights, webinars, blogs and videos
Our platform Discover the cell identity coding platform that powers our ioCells