APP-KM670-671NL-Het_DAPI_MAP2

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.

per vial

For academic discounts, sample requests or bulk pricing inquiries, contact us

Benchtop benefits

In vitro Alzheimer's disease cell model

Disease related phenotype

Overall increase in the production of amyloid beta peptides compared to the wild type control, measured by immunoassay.

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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.

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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.

Technical data

Disease related phenotype

Increased overall production of A𝛽38, A𝛽40 and A𝛽42 shown in ioGlutamatergic Neurons APP KM670/671NL (Swedish), as observed in Alzheimer’s disease
bit.bio glutamatergic neurons Alzheimer's disease model carrying APP Swedish mutation KM670-671NL amyloid-beta quantification

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

bit.bio Alzheimers disease model Swedish mutation ICC staining for glutamatergic neuron  specific markers VGLUT2 MAP2 TUBB3

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

bit.bio Brightfield imaging of iPSC derived neurons Alzheimer's disease model shows glutamatergic neurons morphology

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

bit.bio Alzheimer's disease model Swedish mutation gene expression neuronal markers

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.

Cells arrive ready to plate

iPSC-derived glutamatergic neurons cell culture protocol timeline

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

iPSC-derived excitatory glutamatergic neurons culture in multiple plate formats

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.

Product information

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 10⁶ viable 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 KM670/671NL double point mutation in the APP gene

Applications

Alzheimer's disease research
Drug discovery and development
Disease modelling

Available clones

io1060 | APP KM670/671NL/WT (CLD5)
io1061 | APP KM670/671NL/WT (CLE4)
io1062 | APP KM670/671NL/WT (CLE9)

Product use

ioCells are for research use only

Product resources

Producing 3D Neuronal Microtissues for Preclinical Drug Screening using ioGlutamatergic Neurons Application note
Producing 3D Neuronal Microtissues for Preclinical Drug Screening using ioGlutamatergic Neurons
V1
2024
bit.bio
Inventia
Download
Sartorius application note - Advanced in vitro Modeling of Human iPSC-derived Neuronal Mono- and Co-cultures with Microglia Application note
Sartorius application note - Advanced in vitro Modeling of Human iPSC-derived Neuronal Mono- and Co-cultures with Microglia
Trigg et al.,
Sartorius
2024
Download
ioGlutamatergic Neurons Brochure
ioGlutamatergic Neurons

bit.bio

Download
ioGlutamatergic Neurons Wild Type and related disease models | User Manual User manual
ioGlutamatergic Neurons Wild Type and related disease models | User Manual

V11

bit.bio

2024

Download
Circadian clocks in human cerebral organoids Publication
Circadian clocks in human cerebral organoids

Rzechorzek, et al

bioRxiv

2024

Featuring opti-ox enabled microglia male iPS cell line and opti-ox enabled glutamatergic neurons iPS cell line

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Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease Poster
Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease
Smith, et al. 
bit.bio
2024
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Schizophrenia risk gene ZNF804A controls ribosome localization and synaptogenesis in developing human neurons Publication
Schizophrenia risk gene ZNF804A controls ribosome localization and synaptogenesis in developing human neurons

Deepak P. Srivastava, et al

bioRxiv

2024

Featuring opti-ox enabled glutamatergic neurons iPS cell line

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Generating publishable neuroscience research in 12 weeks with ioGlutamatergic Neurons Case study
Generating publishable neuroscience research in 12 weeks with ioGlutamatergic Neurons

Professor Deepak Srivastava

Professor of Molecular Neuroscience and Group Leader, MRC Centre for Developmental Disorders

King’s College London 

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Running Large-Scale CRISPR Screens in Human Neurons Webinar
Running Large-Scale CRISPR Screens in Human Neurons

Emmanouil Metzakopian | Vice President, Research and Development | bit.bio

Javier Conde-Vancells | Director Product Management | bit.bio

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Rewiring of the promoter-enhancer interactome and regulatory landscape in glioblastoma orchestrates gene expression underlying neurogliomal synaptic communication Publication
Rewiring of the promoter-enhancer interactome and regulatory landscape in glioblastoma orchestrates gene expression underlying neurogliomal synaptic communication

Chakraborty et al
Nature Communications
2023

Featuring ioGlutamatergic Neurons

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Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming Webinar
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming

Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | bit.bio

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Industrialising Cellular Reprogramming: Leveraging opti-ox Technology to Manufacture Human Cells with Unprecedented Consistency Talk
Industrialising Cellular Reprogramming: Leveraging opti-ox Technology to Manufacture Human Cells with Unprecedented Consistency

Innovation showcase talk at ISSCR

Marius Wernig MD, PhD | Stanford 

Mark Kotter, MD, PhD | bit.bio

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Modelling neurodegeneration: Human isogenic system to study FTD & ALS Poster
Modelling neurodegeneration: Human isogenic system to study FTD & ALS

Oosterveen, et al

bit.bio & Charles River Laboratories

2023

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HNRNPH1 regulates the neuroprotective cold‐shock protein RBM3 expression through poison exon exclusion Publication
HNRNPH1 regulates the neuroprotective cold‐shock protein RBM3 expression through poison exon exclusion

Qiaojin Lin et al

The EMBO Journal

2023

Featuring opti-ox powered hiPSC-derived glutamatergic neurons with constitutive expression of Cas9

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Rethinking Developmental Biology With Cellular Reprogramming Webinar
Rethinking Developmental Biology With Cellular Reprogramming

Mark Kotter | CEO and founder | bit.bio

Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology |  Stanford University

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Precision Cellular Reprogramming for Scalable and Consistent Human Neurodegenerative Disease Models Talk
Precision Cellular Reprogramming for Scalable and Consistent Human Neurodegenerative Disease Models

Madeleine Garrett | Field Application Specialist | bit.bio

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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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Wild Type and Isogenic Disease Model cells: A true comparison

Further your disease research by pairing our wild type cells with isogenic disease models.

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Related pages

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