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Description
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 |
| Cell type | iPSC-derived sensory neuron progenitors |
| Growth properties | Adherent |
| Shipping conditions | Dry ice |
| Storage conditions | Vapor phase nitrogen |
Frequently Asked Questions
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.
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.
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 neuron culture has a lot of cell death since I have treated with mitomycin C
Expect significant cell death post mitomycin C treatment. The effect of mitomycin C becomes very evident 4 -5 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 according to the User Guide throughout the differentiation process you will have high-quality networked sensory neurons 2 weeks after thawing.
What percentage of sensory neurons would express TRPV1 in my culture?
At least 70% of terminally differentiated sensory neurons express TRPV1 as validated with ICC and MEA with sensory neurons treated with Capsaicin.
How long can iPSC-derived sensory neurons be kept in culture?
We suggest that you carry out your experiments with the sensory neurons between day 21 and day 29