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Human iPSC-Derived Neural Stem Cells - Huntington's Disease Patient (CAG50)

Huntington's Disease-specific iPSC-derived Neural Stem Cells (CAG 50); > 1.5 million cells per vial.

Can be used with striatal neuron medium kit (ax0333) to generate disease-relevant medium spiny neurons.
Product Code: ax0211-kit Categories: , , .

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Description

Model Huntington's Disease in a Dish

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

  • • Human cells derived from diagnosed Huntington's Disease patients
  • • Available in industrial quantity
  • • High purity neural stem cell population
  • • Ready-made and easy to differentiate to either cerebral cortical or striatal neuron lineages
  • • Robust enough to undergo genetic manipulation
  • • Capable of differentiating to a spectrum of excitatory and inhibitory neurons
  • • Remain viable in culture for months to facilitate reproducible and long term studies

Applications

  • • Studying Huntington's Disease pathogenesis at the cellular level
  • • Studying neural network formation and signal transmission in Huntington's Disease neural culture
  • • Neurotoxicity testing
  • • Preclinical drug efficacy testing
  • • Co-culture studies with astrocytes


Odoo image and text block

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.

Odoo image and text block

Huntington's Disease Neural Stem Cells Express Neural Stem Cell Markers HD NSCs were fixed after 7 days in culture and stained using our immunocytochemisty 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).



Product Specification

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
Kit components 1 vial of Neural Stem Cells (≥1.5 million cells) and 1 bottle of Neural Plating-XF Medium (30 mL)
Cell type iPSC-derived neural stem cells
Growth properties Adherent
Disease information CAG repeat expansion 50
Shipping conditions Dry ice
Storage conditions Liquid nitrogen

Frequently Asked Questions

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.

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 do not recommend using coating reagents whcih we have not tested with our cells. They might result in poor quality cultures and low adherence. Our SureBond, SureBond-XF and SureBond+ReadySet have been optimized to complement our cells and provide consistent results.

We do not recommend re-freeze the NSCs. Axol cannot guarantee the viability of the iPSC-derived NSCs and functionality of the neurons derived after re-freezing.

At day 21, spontaneous synaptic activities should be detected, and day 35 synchronised burst firing should occur.

Technical Resources

References

  • Wevers NR, van Vught R, Wilschut KJ, et al., High-throughput compound evaluation on 3D networks of neurons and glia in a microfluidic platform. Scientific Reports (2016)