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Description
Axol Human iPSC-Derived Sensory Neuron Progenitors derived from a healthy male donor. Axol's induced pluripotent stem cells (iPSCs) were differentiated to sensory neuron progenitors using small molecule inhibitors. For endpoint assays on plastic, SureBond-XF should be used for coating cell culture surfaces. For endpoint assays on glass, SureBond+ReadySet coating reagent should be used. After thawing and plating, the progenitors need to be growth arrested and then should be matured for approximately 6 weeks prior to endpoint assays. The expression of Nav1.7 should occur at around 4-5 weeks of maturation and Nav1.8 is expressed after approximately 6-8 weeks of maturation in Sensory Neuron Maintenance Medium.
Product Specification
| Starting material | Cord blood CD34+ cells |
| Donor gender | Male |
| Donor age at sampling | Newborn |
| HLA serotype | A29 A68, B38(Bw4) B44(Bw4), Cw8 Cw12 |
| Karyotype | Normal |
| Reprogramming method | Episomal vector |
| Induction method | Monolayer & chemically defined medium |
| Genetic modification | None |
| Size | 500,000 cells |
| Kit components | 1 vial of progenitors (>500,000 cells) and 1 bottle of Neural Plating-XF Medium (30 mL) |
| Cell type | iPSC-derived sensory neuron progenitors |
| Growth properties | Adherent |
| Shipping conditions | Dry ice |
| Storage conditions | vapour phase nitrogen |
Frequently Asked Questions
I have plated the cells and saw some flat cells in the background of the neuronal culture, is this normal?
Yes, it is normal. The mitomycin treatment step will eliminate the flat cells.
My sensory culture has a lot of cell death since I have treated with mitomycin
Expect significant cell death post mitomycin C treatment. Mitomycin's effect becomes very evident a couple of days after treatment and this is fine. Don't panic if you see residual mitomycin c induced cell death even 8-10 days post-treatment. If your sensory neuron culture is maintained in a healthy way throughout the differentiation process you will have high-quality networked sensory neurons 2 weeks after thawing.
Do I have to treat the differentiating sensory progenitors with mitomycin C?
Yes, this is a necessary purification step. If you do not treat the cells with mitomycin C, then the flat non-sensory neuron cells would take over the culture.
A sheet of neurons is peeling from the corners , is this normal and can I stop it?
Please make sure you change medium gently and avoid adding the medium from one side of the wells throughout the 5-6 weeks of culture. If the cells start to peel from the corners, it can be repaired by adding Surebond (ax0041) into your standard feeding media. Usually, we use 120 uL Surebond in 12 mL medium for a few days until the layer re-attach. This can be applied no matter what coating has been used.
Which coating shall I use for sensory neuron progenitor differentiation?
They work best on Readyset + Surebond (our strongest coating set). They are OK on Surebond-XF but mature cells tend to tug at each other pulling the layer apart, which surebond-XF can not stop or prevent.
How often do I have to change maintenance medium for sensory neurons?
For terminally differentiated cultures of sensory neurons, you can change medium every 2 to 3 days.
Shall I Filtration Sterilize the medium after Mitomycin C addition before treating the cells?
Yes.
Can I expand the iPSC-derived sensory neuron progenitors?
No, these cells would need to be treated with mitomycin C at day 3 which would halt any remaining proliferative potential.
What percentage of sensory neurons would express TRPV1 in my culture?
After 5 weeks of differentiation, 70-80% of terminally differentiated sensory neurons express TRPV1 as validated with ICC.
How long can iPSC-derived sensory neuron progenitors be kept in culture?
Sensory neurons can be kept in continuous culture for over 10 months. Special culture method is NOT required. The standard culture protocol and Sensory neuron maintenance medium can be used for supporting the long term culture.