Huntington’s Disease-specific iPSC-derived Neural Stem Cells (CAG 50); > 1.5 million cells per vial.
We have applied our innovative technology to develop human neural stem cells directly differentiated from Huntington’s Disease iPS cells. Our cells allow Huntington’s Disease researchers to build their own human cellular models in a disease-relevant genetic and tissue context.
Our ready-to-use kits allow you to concentrate on your experiments rather than spending valuable time and resources generating Huntington’s Disease NSCs in your lab.
HD NSCs are ready made and easy to differentiate. By using our striatal neuron medium, they are able to differentiate to medium spiny neurons expressing key markers, such as DARPP32, CALBINDIN, GABA, CTIP2 and MAP2. By using our cerebral cortical differentiation medium kits, they are capable of differentiating into a spectrum of cortical excitatory and inhibitory neurons and remain viable in culture for months, facilitating reproducible and long-term studies.
Whether you want to study Huntington’s Disease pathogenesis at the cellular level, perform neurotoxicity testing or pre-clinical drug screening, we believe that HD NSCs, alongside our optimized culture reagents provide the perfect culture system for your research success.
Advantages of Axol HD NSCs
Applications
Phase Contrast Image of Huntington’s Disease Human Neural Stem Cells The image was taken at Day 7 in vitro. In the centre of the image you can see HD NSCs in a neural rosette formation. Neurite outgrowth can also be observed.
Huntington’s Disease Neural Stem Cells Express Neural Stem Cell Markers HD NSCs were fixed after 7 days in culture and stained using our protocol. The neural stem cells were counterstained with DAPI (Blue) to visualise the nuclei. The cells can be seen to express neural stem cell markers Ki67 (Green) and Nestin (Red).
| Starting material | Fibroblasts |
| Donor gender | Female |
| Donor age at sampling | 48 yrs |
| Donor age of onset | 45 yrs |
| Reprogramming method | Retroviral |
| Induction method | Monolayer & chemically defined medium |
| Size | ≥1.5 million cells |
| Cell type | iPSC-derived neural stem cells |
| Growth properties | Adherent |
| Disease information | CAG repeat expansion 50 |
| Shipping conditions | Dry ice |
| Storage conditions | Liquid nitrogen |
Plate the cells on Readyset + Surebond (ax0052)
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.
We only recommend using SureBond-XF (Ax0053) in combination with Poly-D-Lysine (Sigma Aldrich, P7405).
We do not recommend expansion or re-freezing of the the NSCs. Axol cannot guarantee the viability of the iPSC-derived NSCs and functionality of the neurons derived after re-freezing or expansion.
At day 21, spontaneous synaptic activities should be detected, and day 35 synchronised burst firing should occur.
ReadySet: 12 mL SureBond: 90 uL Striatal Neuron Medium: 85 mL Below we laid out the calculations: Assuming you’d like to plate the cells at our recommended density – 35,000 cells per square centimetre, you would have 43 square centimetres (or 4.5 wells of a 6-well plate) of cells. Coating: For ReadySet coating, we recommend 250 uL per square centimetre. So, you’d need 11 mL of ReadySet per vial of cells (250 uL x 43 cm2) For SureBond top coating, we recommend adding 120 uL in 6 mL PBS; and then coat 100 uL of diluted SureBond per square centimetre. So, you’d need at least 86 uL of SureBond (in 4.3 mL of PBS) per vial of cells. Medium: Assuming you’d use 6-well plates to start with your experiments, each well would require 2 mL medium. For plating 1 vial of cells (19 mL total): Spin down: 10 mL Then, 2 mL for each well x 4.5 wells = 9 mL Medium A for 1 vial of the cells (40 mL total): Day 1 – complete medium change – 2 mL for each well x 4.5 wells = 9 mL Day 4, 7, 10, 13 and 16 – 2/3 medium change (1.33 mL) – 6.7 mL total for each well x 4.5 wells = 30.2 mL Medium B for 1 vial of the cells (31 mL total): Day 19, 22, 25, 28 and 31 – 2/3 medium change – 6.7 mL x 4.5 wells = 30.2 mL Hence, you’d need 81 mL of medium per vial of cells to differentiate and mature the cells to Day 33 for assaying.
