cat no | io1072
A rapidly maturing, physiologically relevant, functional system for investigating the role of the heterozygous PSEN1 M146L mutation in early-onset Alzheimer's disease (AD). This in vitro disease cell model recapitulates an increase in the amyloid beta peptide ratios observed in AD patients.
ioGlutamatergic Neurons PSEN1 M146L/WT are opti‑ox deterministically programmed human glutamatergic neurons carrying a genetically engineered heterozygous M146L mutation in the PSEN1 gene encoding presenilin 1 protein.
This disease model is part of an Alzheimer's disease panel of human iPSC-derived cells that can be incorporated into translational research and drug discovery workflows. Two additional clones for the PSEN1 M146L/WT mutation are available for scientists who wish to repeat their experiments in multiple independent clones, please enquire. All disease models are genetically matched to the wild-type control, ioGlutamatergic Neurons. Additional mutations in the AD panel include homozygous PSEN1 M146L, heterozygous and homozygous APP KM670/671NL and APP V717I.
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.
per vial
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@ÎÞÓǶÌÊÓƵ.
Disease-related phenotype
Increased ratio of Að›½42:40 and Að›½42:38 peptides compared to the genetically matched control, measured by immunoassay.
Make True Comparisons
Pair the Alzheimer's disease model cells with the genetically matched wild-type ioGlutamatergic Neurons to investigate the impact of the PSEN1 missense mutation on early-onset AD.
Quick
The disease model cells and wild-type control are experiment ready as early as 2 days post revival, and form structural neuronal networks at 11 days.
PSEN1 M146L disease models recapitulate an increase in amyloid beta peptide ratios observed in Alzheimer's disease patients
ioGlutamatergic Neurons PSEN1 M146L disease model cells show an increase in the ratio of Að›½42:40 and Að›½42:38 compared to the wild type, genetically matched control.
ioGlutamatergic Neurons wild type (WT) and PSEN1 M146L heterozygous (HET) clones CL8 (io1072S), CL60 and CL57, and homozygous (HOM) clones CL15 (io1069S), CL19 and CL57, 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; data shown for day 30 only.
Levels of Að›½38, Að›½40 and Að›½42 peptides were quantified using the V-PLEX A𛽠Peptide Panel 1 (6E10) Kit (). Data were obtained from two independent experiments and are shown as mean ± SEM. Data were analysed statistically using one-way ANOVA with Tukey’s post-hoc analysis. * p<0.05 ***p<0.001 **** p<0.0001.
Expression levels for specific genes of interest in the disease model products io1072 (CL8, het) and io1069 (CL15, hom) and the additional clones (CL60, CL71, CL19, CL57) can be requested by contacting technical@ÎÞÓǶÌÊÓƵ
ioGlutamatergic Neurons PSEN1 M146L/WT express neuron-specific markers comparably to the wild type 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 PSEN1 M146L/WT compared to the genetically matched control. 100X magnification.
ioGlutamatergic Neurons PSEN1 M146L/WT form structural neuronal networks by day 11
ioGlutamatergic Neurons PSEN1 M146L/WT 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 PSEN1 M146L/WT demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following deterministic programming
Gene expression analysis demonstrates that ioGlutamatergic Neurons PSEN1 M146L/WT and wild-type ioGlutamatergic Neurons (WT Control) lack the expression of pluripotency markers (NANOG and OCT4) at day 11, while 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.
ioGlutamatergic Neurons PSEN1 M146L/WT 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.
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.
Starting material
Human iPSC line
Karyotype
Normal (46, XY)
Seeding compatibility
6, 12, 24, 96 & 384 well plates
Shipping info
Dry ice
Donor
Caucasian adult male, age 55-60 years old (skin fibroblast)
Vial size
Small: >1 x 10ⶠviable cells
Quality control
Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)
Differentiation method
opti-ox deterministic cell programming
Recommended seeding density
30,000 cells/cm²
User storage
LN2 or -150°C
Format
Cryopreserved cells
Genetic modification
Heterozygous M146L missense mutation in the PSEN1 gene
Applications
Alzheimer's disease research
Drug discovery and development
Disease modelling
Available clones
io1072 | PSEN1 M146L/WT (CL8)
io1073 | PSEN1 M146L/WT (CL60)
io1074 | PSEN1 M146L/WT (CL71)
Product use
ioCells are for research use only
ÎÞÓǶÌÊÓƵ
V11
ÎÞÓǶÌÊÓƵ
2024
Professor Deepak Srivastava
Professor of Molecular Neuroscience and Group Leader, MRC Centre for Developmental Disorders
King’s College London
Emmanouil Metzakopian | Vice President, Research and Development | ÎÞÓǶÌÊÓƵ
Javier Conde-Vancells | Director Product Management | ÎÞÓǶÌÊÓƵ
Chakraborty et al
Nature Communications
2023
Featuring ioGlutamatergic Neurons
Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | ÎÞÓǶÌÊÓƵ
Innovation showcase talk at ISSCR
Marius Wernig MD, PhD | Stanford
Mark Kotter, MD, PhD | ÎÞÓǶÌÊÓƵ
Oosterveen, et al
ÎÞÓǶÌÊÓƵ & Charles River Laboratories
2023
Qiaojin Lin et al
The EMBO Journal
2023
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