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axoCells™ neural plating media, 30 ml
axoCells™ neural plating media, 30 ml is optimized for culture of neural cells for neuroscience research. Suitable for use with axoCells cortical neurons, inhibitory interneurons and sensory neurons.
• Optimized to support maximum post-thaw recovery of neural stem cells
Specifications
Volume 30ml
Shipping conditions Dry ice
Storage conditions -80°C
Description
axoCells™ neural plating media, 30 ml has been optimized for maximum post thaw recovery of neural cells and is used in Axol’s cortical excitatory neuron, inhibitory interneuron and sensory neuron protocols. It is available as a standalone product or as part of the axoCells Human iPSC-Derived Cortical Neuron Cells, Media, Supplements and Coating kits (ax5113, ax5115) and axoCells Human iPSC-Derived Sensory Neuron Cells, Media, Supplement and Coating kit (ax0157).
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Additional Information
Cortical Excitatory Neurons
Applications
Our axoCellsTM cortical excitatory neuron progenitors are suitable for investigating Alzheimer’s disease and for screening candidate therapies for this disease.
Healthy control lines are also available and can be used with high–content imaging, MEA, microfluidic and organ-on-chip (OOC) devices to measure structural and functional responses in neuronal models.
Protocols
Fully optimized cell culture reagents are available for the differentiation of progenitors into mature cortical excitatory neurons within 14 days. In addition, single media protocols, enabling co- and tri-culture of our cortical excitatory neurons with inhibitory interneurons, astrocytes and microglia provide convenient and fast culture solutions to better model human diseases in vitro.
Quality
We maintain or exceed industry-level quality with our ISO:9001-accredited production facility, guided by our rigorous quality control procedures and decades of scientific experience.
All cells come with a full Certificate of Analysis and are officially certified by HPSCreg®, to ensure ethical and biological conformity for your peace of mind.
Licenses & Consent
Axol has obtained all relevant licenses for reprogramming donor samples into iPSCs and differentiating these into cortical excitatory neurons for commercial use.
Patient samples used to create these neurons have been ethically sourced and consented for research and commercial use. All cells come with a full Certificate of Analysis and are officially certified by HPSCreg®, to ensure ethical and biological conformity for your peace of mind.
Publications
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498259/
Garcia-Leon JA, Vitorica J, & Gutierrez A (2019). Use of human pluripotent stem cell-derived cells for neurodegenerative disease modeling and drug screening platform. Future Medicinal Chemistry, 11(11), 1305–1322. doi: 10.4155/fmc-2018-0520 [PubMed] [CrossRef] [Google Scholar]
Media and Reagents
Using NeurOne cortical neuron supplement, mature cerebral cortical excitatory neurons can be generated in 20 days. We also supply neural plating media, maintenance media and SureBond –XF coating as standalone items or as a kit.
Cortical Inhibitory Interneurons
Applications
Our axoCellsTM cortical inhibitory interneurons can be used in co-culture with axoCellsTM cortical excitatory neurons to investigate the connections in the brain and also monitor the effect of inhibitory neurons on the electrical activity of these excitatory neurons, mimicking conditions such as epilepsy. In order to create more physiologically relevant models, the inclusion of microglia and/or astrocytes is also possible.
Protocols
Mono- and co-culture protocols for interneurons alone or with cortical excitatory neurons are available. In addition, we have protocols for tri-culture with astrocytes or microglia using one media for all cell types.
Quality
As an Early Access Product, these cells will not have been manufactured at our ISO accredited facility and final protocols, documentation and literature may still be development.
We welcome feedback on these products from the scientific community. These products are available to purchase at a discounted package price and will be supplied with a feedback form.
Licenses & Consent
Axol has obtained all relevant licenses for reprogramming donor samples into iPSCs and differentiating these into cortical inhibitory interneurons for commercial use.
Patient samples used to create these neurons have been ethically sourced and consented for research and commercial use. These early access cells come with a cellular analysis datasheet and are officially certified by HPSCreg®, to ensure ethical and biological conformity for your peace of mind.
Publications
Cruz-Santos M, Cardo LF, Li M. A Novel LHX6 Reporter Cell Line for Tracking Human iPSC-Derived Cortical Interneurons. Cells. 2022 Mar 1;11(5):853. doi: 10.3390/cells11050853.
PMID: 35269475; PMCID: PMC8909769.
Dubonyte, U., Asenjo-Martinez, A., Werge, T. et al. Current advancements of modelling schizophrenia using patient-derived induced pluripotent stem cells. acta neuropathol commun 10, 183 (2022). https://doi.org/10.1186/s40478-022-01460-2
Ni, P., Noh, H., Park, GH. et al. iPSC-derived homogeneous populations of developing schizophrenia cortical interneurons have compromised mitochondrial function. Mol Psychiatry 25, 2873–2888 (2020). https://doi.org/10.1038/s41380-019-0423-3
Media & Reagents
We supply optimized media and reagents for axoCells cortical inhibitory interneuron culture, whether alone or in co- or tri-culture with other neurons or glia cells. Neural Maintenance Media (ax0031), NeurOne Cortical Neuron Supplement (ax0679), Neural Plating Media (ax0033) and SureBond-XF (ax0053) coating are all available as standalone products or the culture media and coating are available in a kit (Neural Stem Cell Media & Reagent Bundle ax0105) containing ax0674, ax0031 and ax0053.
Sensory Neurons
Phase contrast
Phase contrast images show the differentiation and maturation of axoCells Sensory Neuron Progenitors over two weeks after thawing and treatment with mitomycin C.
Phase contrast images of axoCells Sensory Neuron Progenitors The cells were plated on SureBond-XF in Neural Plating Medium. The cells were then treated with mitomycin C two days after thawing and cultured in the supplemented Sensory Neuron Maintenance Medium. (axoCells sensory neurons should be cultured for a minimum of 3 weeks prior to performing endpoint assays.)
Immunocytochemistry
Immunocytochemistry (ICC) images of axoCells sensory neurons show expression of the key marker TUJ-1, indicative of neuronal development, and the nociceptive ion channels TrpV1, Nav1.7 and Nav1.8, responsible for the generation and maintenance of abnormal neuronal electrogenesis and hyperexcitability in the development of pathological pain.
ICC of axoCells Sensory Neuron Progenitors The cells were plated on SureBond-XF in Neural Plating Medium. The cells were then treated with mitomycin C two days after thawing and cultured in the supplemented Sensory Neuron Maintenance Medium. (axoCells sensory neurons should be cultured for a minimum of 3 weeks prior to performing endpoint assays.)
Culturing axoCells sensory neurons with Maximizer supplement speeds up maturation. On average we see increases of 30-45% of neurons expressing TRPv1 and Nav1.7 upon treatment with Maximizer.
Sodium ion channel expression
Sodium channel RNA expression analysis by cDNA PCR
axoCells Sensory Neuron Progenitors show RNA expression of all three voltage-gated sodium ion channels, Nav1.7, Nav1.8 and Nav1.9.
cDNA from axoCells Sensory Neurons 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 axoCells sensory neurons. SCN5a (237 bp, hNav1.5) was included as a negative control. Data provided by Dr Edward Emery (University College London).
Capsaicin and menthol responses
axoCells sensory neurons were challenged with capsaicin and menthol at 22 and 27 days respectively. As early as 22 days, 90% of the neurons responded to capsaicin challenge with increased spike measurements on a multi electrode array (MEA) platform.
Initial menthol application caused an increase in spike firing, demonstrating appropriate functional activity of the sensory neurons, but repeated application was seen to trigger a decrease in spike firing or tachyphylaxis (i.e. desensitization).
Dose responses for both chemicals were observed, demonstrating how the axoCells sensory neurons could be used as advanced in vitro models of pain, sensation, and peripheral nervous system disorders for drug discovery and cosmetic toxicity testing.
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 axoCells sensory neurons.
Electrophysiological characterization of axoCells sensory neurons using patch clamp. A) Phase contrast image of axoCells sensory neurons; 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).
Mustard Oil and ATP treatment
Mustard Oil treatment of axoCells sensory neurons elicited a significant increase in the number of spikes on MEA, indicating the contribution this agent plays to the pain response. Mustard oil is an agonist of the TRPA1 (transient receptor potential ankyrin 1) receptor.
ATP treatment of axoCells sensory neurons also elicited a significant increase in the number of spikes on MEA. ATP opens ligand-gated ion channels (P2X receptors) in sensory neurons.
Thermoception
A change in the firing pattern of axoCells sensory neurons was seen in response to temperature change. Increased firing on MEA was seen in axoCells sensory neurons in response to heating and decreased firing in response to cooling. This demonstrates thermoception, an in vivo function, and shows axoCells sensory neurons to be functionally relevant cells.
Paclitaxel
Treatment of axoCells sensory neurons with the chemotherapy reagent paclitaxel demonstrates reduced neurite length (axotomy) and the functional relevance of the sensory neurons. This provides a model for both acute insult and chronic cytotoxicity.
Response of axoCells sensory neurons to paclitaxel, a chemotherapy reagent, applied at double arrow (washoff at single arow).
Applications
Our axoCellsTM sensory neurons have been specifically developed for use in microfluidics systems, organ-on-chip devices and for in vitro disease modelling.
We have a community of top ten biopharma and research institutes who use our sensory neurons to build powerful in vitro models, investigating disorders of the peripheral nervous system, chronic pain and analgesic drug screening.
Protocols
We offer a fully optimized cell culture system including tailored Sensory Maintenance Medium to promote the viability and maturation of sensory neurons for endpoint assays on glass or plastic.
Quality
We maintain or exceed industry-level quality with our ISO:9001-accredited production facility, guided by our rigorous quality control procedures and decades of scientific experience.
All cells come with a full Certificate of Analysis and are officially certified by HPSCreg®, to ensure ethical and biological conformity for your peace of mind.
Licenses & Consent
Axol has obtained all relevant licenses for reprogramming donor samples into iPSCs and differentiating these into sensory neurons for commercial use.
Patient samples used to create these neurons have been ethically sourced and consented for research and commercial use. All cells come with a full Certificate of Analysis and are officially certified by HPSCreg®, to ensure ethical and biological conformity for your peace of mind.
Publications
The RNA-Binding Protein HuR Is Integral to the Function of Nociceptors in Mice and Humans- Kunder N,et al. 2022. J Neurosci. doi: 10.1523/JNEUROSCI.1630-22.2022. Epub 2022 Oct 21. PMID: 36270801; PMCID: PMC9761683.
Macrophage epigenetic memories of early life injury drive neonatal nociceptive priming- Dourson AJ et al. bioRxiv [Preprint]. doi: 10.1101/2023.02.13.528015.
SARS-CoV-2 papain-like protease activates nociceptors to drive sneeze and pain.- Mali et al 2024. bioRxiv [Preprint]. 2024 doi: 10.1101/2024.01.10.575114.
Emerging Neurotechnology for Antinoceptive Mechanisms and Therapeutics Discovery – Black et al 2018 Biosensors and Bioelectronic https://doi.org/10.1016/j.bios.2018.11.015
A Microfluidic Approach to Investigate Changes in Functional Properties of Nociceptive Axons Underlying Inflammatory Pain States – Natasha Rangwani 2018. PhD Thesis
Human stem cell derived sensory neurons are positioned to support varicella zoster virus latency – Sadaoka et al. 2020 BioRxiv https://doi.org/10.1101/2020.01.24.919290
Varicella-zoster virus VLT-ORF63 fusion transcript induces broad viral gene expression during reactivation from neuronal latency – Ouwendijk et al. 2020 Nature Communications https://doi.org/10.1038/s41467-020-20031-4
Mechanistic insights into the pathogenesis of microtubule-targeting agent-induced peripheral neuropathy from pharmacogenetic and functional studies – Chua et al. 2021 BCPT https://doi.org/10.1111/bcpt.13654
Considerations for a Reliable In Vitro Model of Chemotherapy-Induced Peripheral Neuropathy – Eldridge et al. 2021 Toxics https://doi.org/10.3390/toxics9110300
In Vitro Pain Assay Using Human iPSC-Derived Sensory Neurons and Microelectrode Array – Odawara et al. 2022 Toxicological Sciences https://doi.org/10.1093/toxsci/kfac045
Global analyses of mRNA expression in human sensory neurons reveals eIF5A as a conserved target for inflammatory pain – Chase et al. 2022 FASEB J. https://doi.org/10.1096%2Ffj.202101933RR
Use of a human induced pluripotent stem cell-derived dorsal root ganglion neurone model to study analgesics in vitro: proof of principle using lidocaine – Smulders et al. 2022 British Journal of Anaesthesia https://doi.org/10.1016/j.bja.2022.09.009
Chow SYA, et al. 2022. Human sensory neurons modulate melanocytes through secretion of RGMB. Cell Rep. 2022 Sep 20;40(12):111366. doi: 10.1016/j.celrep.2022.111366. PMID: 36130522.
Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury – Nishijima et al. 2023 Research Square https://doi.org/10.21203/rs.3.rs-3503996/v1
Scalable generation of sensory neurons from human pluripotent stem cells – Deng et al. 2023 Stem Cell Reports https://doi.org/10.1016/j.stemcr.2023.03.006
New human in vitro co-culture model of keratinocytes and sensory neurons like cells releasing substance P with an evaluation of the expression of ZIKV entry receptors: A potent opportunity to test Zika virus entry and to study Zika virus’ infection in neurons? – Bocciarelli et al. 2023 Experimental Dermatology https://doi.org/10.1111/exd.14870
Deng T, et al. 2023. Scalable generation of sensory neurons from human pluripotent stem cells. Stem Cell Reports. 2023 Apr 11;18(4):1030-1047. doi: 10.1016/j.stemcr.2023.03.006. PMCID: PMC10147831.
Media & Reagents
Ensure high-quality, mature sensory neurons in just 21 days with our fully optimized media Sensory Neuron Maximizer kit (ax0158) and Maturation Maximizer media supplement (ax0058). We also supply cells, all growth factors and coating materials as a kit Human iPSC-Derived Sensory Neuron Maximizer Kit (ax0157) with cells).
