Human iPSC-Derived Sensory Neuron Progenitors

Axol Human iPSC-Derived Sensory Neuron Progenitors

iPSC-derived sensory neuron progenitors

We’ve used our expertise in neural differentiation to bring you dorsal root ganglion (DRG) neurons derived from induced pluripotent stem cells (iPSCs). Axol Human iPSC-Sensory Neuron Progenitors are derived from integration-free iPSCs of a healthy male donor and have been differentiated to neurons using small molecule inhibitors. We also offer a fully optimized cell culture system including tailored Sensory Neuron Maintenance Medium and coating reagents to promote the viability and maturation of sensory neurons for endpoint assays on glass or plastic.

Our iPSC-derived sensory neurons express several voltage-gated sodium ion channels and transient receptor potential (TRP) ion channels that play a key role in nociception. These include sodium ion channels Nav1.7 and the DRG-specific, TTX-resistant channels, Nav1.8 and Nav1.9 as well as the temperature-sensitive, TRPV1 and TRPM8, and TRPA1, a sensor of pungency, bitterness and cold.

Axol iPSC-Derived Sensory Neuron Progenitors are available in large batch sizes for reliable and consistent results in high-throughput screening assays. The cells are also suitable for investigating disorders of the peripheral nervous system and chronic pain as well as drug targets for pain relief.


iPSC-derived sensory neuron progenitors

Cat. No. Product Name Starting Material Quantity Price
ax0055 Human iPSC-Derived Sensory Neuron Progenitors (Male) Cord Blood CD34+ Cells 500,000 cells & Neural Plating-XF Medium $560

Sensory neuron culture media and reagents

Cat. No. Product Name Product Description Quantity Price
ax0060 Sensory Neuron Maintenance Medium Medium for the maturation and maintenance of iPSC-Derived Sensory Neuron Progenitors 250 ml $200
ax0041+/ax0052 SureBond+ReadySet Coating reagent needed for endpoint assays on glass 3 x 120 μL vials & 2 x 10 mL vials $240
ax0041XF/ax0053 SureBond-XF Xeno-free coating reagent needed for endpoint assays on plastic 1 mL $400

Phase contrast

Phase contrast images show the maturation of Axol Human iPSC-Derived Sensory Neuron Progenitors over two weeks after thawing and treatment with mitomycin C.

Phase contrast image of Axol Human iPSC-Derived Sensory Neuron Progenitors

Phase contrast of Axol Human iPSC-Derived Sensory Neuron Progenitors The cells were plated on SureBond-XF in Neural Plating-XF Medium. The cells were then treated with mitomycin C two days after thawing and cultured in Sensory Neuron Maintenance Medium containing growth factors (GDNF, NGF, BDNF and NT-3) for two weeks. (iPSC-sensory neurons should be culture for a minimum of 6 weeks prior to performing endpoint assays.)


Sodium ion channel expression

Sodium channel RNA expression analysis by cDNA PCR

Axol iPSC-Derived Sensory Neuron Progenitors show RNA expression of all three voltage-gated sodium ion channels, Nav1.7, Nav1.8 and Nav1.9.

Sodium ion channel RNA expression in Axol Human iPSC-Derived Sensory Neuron Progenitors

cDNA from iPSC-Derived Sensory Neuron Progenitors cultured for 8 weeks was compared to cDNA from human tissue from the dorsal root ganglion (DRG). PCR analysis (40 cycles; 55oC ) confirmed the mRNA expression of SCN9A (82 bp, hNav1.7), SCN10A (149 bp, hNav1.8) and SCN11A (464 bp, hNav1.9) in Axol iPSC-derived sensory neurons. SCN5a (237 bp, hNav1.5) was included as a negative control. Data provided by Dr Edward Emery (University College London).

Nav1.7 immunocytochemistry

Axol Human iPSC-Derived Sensory Neuron Progenitors express Nav1.7, a key sodium ion channel in DRG sensory neurons that plays an important role in peripheral pain sensation.

Nav1.7 expression in Axol Human iPSC-Derived Sensory Neuron Progenitors

Axol Human iPSC-Derived Sensory Neuron Progenitors express the nociceptive voltage-gated sodium ion channel Nav1.7 after 63 days culture in Sensory Neuron Maintenance Medium on a multi-electrode array (MEA) (Alpha MED Scientific). Nav1.7 (red), β-tubulin Ⅲ (green), Hoechst (blue). Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

Nav1.7 and Nav1.8 immunocytochemistry

Mature iPSC-derived sensory neurons express the sodium ion channels, Nav1.7 and Nav1.8, and the maturation marker, doublecortin (DCX) after 6 weeks in culture.

Axol iPSC-derived sensory neurons express Nav1.7, Nav1.8 and doublecorticon

Axol Human iPSC-Derived Sensory Neuron Progenitors show positive staining for the nociceptive voltage-gated sodium ion channels A) Nav1.7 (green), Nav1.8 (red), DAPI (blue) after 38 days in culture and B) Nav1.8 (red) and doublecortin (green) after 42 days in culture.

Sodium channel–mediated response to potassium chloride

Axol iPSC-Derived Sensory Neuron Progenitors have been shown to elicit a calcium response to the treatment of KCl at the soma signifying the presence of sodium channels.

Somal response to 15 mM potassium chloride in iPSC-derived sensory neurons
Average peak response to potassium cholride in iPSC-derived sensory neurons

TTX-resistant Nav1.8 and Nav1.9

Nociceptive sensory neurons are unique in that they contain voltage-gated inward current sodium channels (Nav 1.8 and Nav 1.9) that are resistant to tetrodotoxin (TTX). The presence of these TTX-resistant ion channels was confirmed in Axol iPSC-derived sensory neurons.

Electrophysiological characterization of Axol Human iPSC-Derived Sensory Neuron Progenitors

Electrophysiological characterization of Axol Human iPSC-Derived Sensory Neuron Progenitors using patch clamp after 56 days in culture A) Phase contrast image of iPSC-derived sensory neurons at 8 weeks; B) Example of a sodium-current elicited by a voltage step from -100 mV to -25 mV in the presence of tetrodotoxin (0.5 mM); C) Current-voltage plot of averaged Na-currents recorded from iPSC-derived sensory neurons in the presence of TTX (n=9). Data provided by Dr Edward Emery (University College London).


Transient receptor potential ion channel expression

TRPA1 immunocytochemistry

Axol Human iPSC-Derived Sensory Neuron Progenitors express TRPA1, the ion channel involved in sensing cold, pungency and bitterness.

Axol Human iPSC-Derived Sensory Neuron Progenitors express the TRPA1 ion channel

Axol Human iPSC-Derived Sensory Neuron Progenitors express the TRPA1 ion channel after 63 days culture in Sensory Neuron Maintenance Medium on a multi-electrode array (MEA) (Alpha MED Scientific). TRPA1 (purple), β-tubulin Ⅲ (green), Hoechst (blue). Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

TRPA1–mediated response to allyl isiothiocyanate 

Axol iPSC-derived sensory neurons show a short burst of firing after the application of allyl isiothiocyanate (AIT), which indicates the presence of TRPA1 channels.

Axol human iPSC-derived sensory neurons show a short burst of firing after the application of 100 μM allyl isothiocyanate

Axol human iPSC-derived sensory neurons show a short burst of firing after the application of 100 μM allyl isothiocyanate. The cells were cultured at 5.0 × 105 cells/cm2 on 64-channel MED-P515A MEA chips (Alpha MED Scientific) coated with SureBond+ReadySet for 7 weeks to obtain mature sensory neurons. This rastor plot shows the firing frequency of sensory neurons before and after the application of 100 μM allyl isothiocyanate (AIT). A short burst of firing after the application of AIT is observed, as expected. Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

TRPV1 immunocytochemistry

Axol Human iPSC-Derived Sensory Neuron Progenitors express TRPV1, the ion channel involved in sensing heat.

TRPV1 expression in Axol Human iPSC-Derived Sensory Neuron Progenitors

Axol Human iPSC-Derived Sensory Neuron Progenitors express the TRPV1 ion channel after 63 days culture in Sensory Neuron Maintenance Medium on a multi-electrode array (MEA) (Alpha MED Scientific). TRPV1 (yellow), β-tubulin Ⅲ (green), Hoechst (blue). Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

TRPV1–mediated response to capsaicin

Axol iPSC-Derived Sensory Neuron Progenitors have been shown to elicit a calcium response to the treatment of capsaicin at both the soma and the axon. This indicates that the iPSC-Derived Sensory Neurons express TRPV1 and can sense heat.

Somal response to 500 nM capsaicin in iPSC-derived sensory neurons
Axonal response to 500 nM capsaicin in iPSC-derived sensory neurons

Axol iPSC-derived sensory neurons have been shown to increase neuronal firing after the application of capsaicin, which indicates the presence of TRPV1 channels.

Axol human iPSC-derived sensory neurons show sustained firing after the application of 100 nM capsaicin

Axol human iPSC-derived sensory neurons show sustained firing after the application of 100 nM capsaicin. The cells were cultured at 5.0 × 105 cells/cm2 on 64-channel MED-P515A MEA chips (Alpha MED Scientific) coated with SureBond+ReadySet for 6 weeks to obtain mature sensory neurons. This rastor plot shows the firing frequency of sensory neurons before and after the application of 100 nM capsaicin. Sustained firing is observed after the application of capsaicin. Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

TRPV1-mediated response to temperature 

Axol iPSC-Derived Sensory Neurons subjected to an increase in temperature at day 14 and 21 results in an increase in neuronal firing. This suggests the expression of TRPV1.

TRPV1–mediated response to temperature in Axol iPSC-derived sensory neurons

Axol human iPSC-derived sensory neurons show an increase in neuronal firing in response to temperature The cells were cultured at 5.0 × 105 cells/cm2 in a 37°C , 5% CO2 atmosphere on 64-channel MED-P515A MEA chips (Alpha MED Scientific) coated with SureBond+ReadySet. Spontaneous and evoked extracellular field potentials were acquired at a sampling rate of 20 kHz/channel and signals were high-pass filtered at 100 Hz. Firing analyses and spike sortings were performed using Mobius software (Alpha MED Scientific) on days 14 (n=218) and 21 (n=510) (Alpha MED Scientific). Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).

TRPM8-mediated response to menthol

Axol iPSC-derived sensory neurons show an increase in firing frequency after the application of menthol, which indicates the presence of TRPM8 channels.

Axol human iPSC-derived sensory neurons show an increase in firing frequency after the application of 100 μM menthol

Axol human iPSC-derived sensory neurons show an increase in firing frequency after the application of 100 μM menthol. The cells were cultured at 5.0 × 105 cells/cm2 on 64-channel MED-P515A MEA chips (Alpha MED Scientific) coated with SureBond+ReadySet for 7 weeks to obtain mature sensory neurons. This rastor plot shows the firing frequency of sensory neurons before and after the application of 100 μM menthol. Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).


Electrical activity

Axol Human iPSC-Derived Sensory Neuron Progenitors display electrical activity.

Electrical activity in Axol Human iPSC-Derived Sensory Neuron Progenitors

Axol Human iPSC-Derived Sensory Neuron Progenitors show spontaneous extracellular field potential on a 64-channel MEA (Alpha MED Scientific) after 33 days in culture. Cells were seeded at 5x105 cells/cm2 and treated with 2.5 µg/ml mitomycin C. Data provided by Prof Ikuro Suzuki (Tohoku Institute of Technology).


Posters 

In vitro pain responses of dorsal root ganglion neurons using multi-electrode arrays

Pain responses in human iPSC-derived sensory neurons using MEA system


Application Note

How to use sensory neurons in MEA-MED64 platform