cat no | io1061
ioGlutamatergic Neurons APP KM670/671NL/WT
Human iPSC-derived Alzheimer's disease model
A rapidly maturing, physiologically relevant, functional system for investigating the role of the APP Swedish mutation in early-onset Alzheimer's disease (AD). This in vitro disease cell model recapitulates an overall increase in the production of amyloid beta peptides, as observed in AD.
ioGlutamatergic Neurons APP KM670/671NL / WT are opti‑ox deterministically programmed glutamatergic neurons carrying a genetically engineered heterozygous double mutation in the APP gene encoding amyloid precursor protein.
Place your order
Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
Benchtop benefits
Disease related phenotype
Overall increase in the production of amyloid beta peptides compared to the wild type control, measured by immunoassay.
Make True Comparisons
Pair the ioDisease Model Cells with the genetically matched wild-type ioGlutamatergic Neurons to investigate the impact of the APP double point mutation on early-onset AD.
Quick
The disease model cells and isogenic control are experiment ready as early as 2 days post revival, and form structural neuronal networks at 11 days.
Disease related phenotype
ioGlutamatergic Neurons APP KM670/671NL Alzheimer's disease model cells show an increase in the overall production of A𝛽38, A𝛽40 and A𝛽42 compared to the wild type, genetically matched control (A), and no change in the ratios of the A𝛽 peptides (B).
- ioGlutamatergic Neurons wild type (WT, io1001) and APP KM670/671NL Hom (CLH12, io1059S) and Het (CLE4, io1061S), were seeded at 30,000 cells/cm2 in 24-well plates and cultured for 30 days according to the user manual. Supernatant was collected at days 10, 20, and 30.
- Levels of A𝛽38, A𝛽40 and A𝛽42 peptides were quantified using the V-PLEX A𝛽 Peptide Panel 1 (6E10) Kit (MSD K15200E-1).
- Concentrations of A𝛽38, A𝛽40, A𝛽42 were normalised to the calculated total number of cells per well.
- Data were obtained from two independent experiments and are shown as mean ± SEM. Data were analysed statistically (at days 20 and 30) using Student’s t-tests comparing each disease model to the wild type.
* p<0.05 ** p<0.01
Highly characterised and defined
ioGlutamatergic Neurons APP KM670/671NL het express neuron-specific markers comparably to the genetically matched control
Immunofluorescent staining on post-revival day 11 demonstrates similar homogenous expression of pan-neuronal proteins MAP2 and TUBB3 (upper panel) and glutamatergic neuron-specific transporter VGLUT2 (lower panel) in ioGlutamatergic Neurons APP KM670/671NL het clones compared to the genetically matched control. 100X magnification.
ioGlutamatergic Neurons APP KM670/671NL het form structural neuronal networks by day 11
ioGlutamatergic Neurons APP KM670/671NL het mature rapidly, show glutamatergic neuron morphology and form structural neuronal networks over 11 days, highly similar to the genetically matched control. Day 1 to 11 post thaw; 100X magnification.
ioGlutamatergic Neurons APP KM670/671NL het demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following deterministic programming
Gene expression analysis demonstrates that ioGlutamatergic Neurons APP KM670/671NL het and wild-type ioGlutamatergic Neurons (WT Control) lack the expression of pluripotency markers (NANOG and OCT4) at day 11, whilst robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic-specific (VGLUT1 and VGLUT2) markers, as well as the glutamate receptor GRIA4. Gene expression levels were assessed by RT-qPCR (data normalised to HMBS; cDNA samples of the parental human iPSC line (hiPSC) were included as reference). Data represents day 11 post-revival samples, n=2 replicates.
Disease-related APP is expressed in ioGlutamatergic Neurons APP KM670/671NL het following deterministic programming
RT-qPCR analysis demonstrates expression of the APP gene in both wild type ioGlutamatergic Neurons (WT Control) and ioGlutamatergic Neurons APP KM670/671NL het at day 11 post-revival. Data normalised to HMBS, n=2 replicates.
Cells arrive ready to plate
ioGlutamatergic Neurons APP KM670/671NL het are delivered in a cryopreserved format and are programmed to mature rapidly upon revival in the recommended media. The protocol for the generation of these cells is a two-phase process: Phase 1, Stabilisation for 4 days; Phase 2, Maintenance, during which the neurons mature. Phases 1 and 2 after revival of cells are carried out by the customer.
Industry leading seeding density
The recommended minimum seeding density is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other similar products on the market. One small vial can plate a minimum of 0.7 x 24-well plate, 1 x 96-well plate, or 1.5 x 384-well plates. This means every vial goes further, enabling more experimental conditions and more repeats, resulting in more confidence in the data.
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Cell culture hacks | human iPSC-derived glutamatergic neurons
Read this blog on glutamatergic neuron cell culture for our top tips on careful handling, cell plating and media changes to achieve success from the outset.
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ioDisease Model Cells
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Wild Type and Isogenic Disease Model cells: A true comparison
Further your disease research by pairing our wild type cells with isogenic disease models.
