APP-KM670-671NL-hom-ICC-DAPI-MAP2

cat no | io1059

ioGlutamatergic Neurons
APP KM670/671NL / KM670/671NL

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 / KM670/671NL are opti‑ox deterministically programmed glutamatergic neurons carrying a genetically engineered homozygous 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.

Glutamatergic neurons Alzheimer's disease in vitro model

Make True Comparisons

Pair the Alzheimer's disease model cells with the genetically matched wild type glutamatergic neurons to investigate the impact of the APP double point mutation on early-onset AD.

Human cortical excitatory neurons culture networks in 11 days

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 Het (CLE4, io1061S) and Hom (CLH12, io1059S), 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 hom 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 TUBB3 and MAP2 (upper panel) and glutamatergic neuron specific transporter VGLUT2 (lower panel) in glutamatergic neurons carrying the Alzheimer's disease related APP KM670/671NL mutation compared to the genetically matched control. 100X magnification.
ioGlutamatergic Neurons APP KM670/671NL / KM670/671NL 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 hom 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 thawing; 100X magnification.
ioGlutamatergic Neurons APP KM670/671NL / KM670/671NL demonstrate gene expression of neuronal and glutamatergic-specific markers following deterministic programming
bit.bio Glutamatergic neuron gene expression in ipsc derived Alzheimer's disease cell model by RT-qPCR
Gene expression analysis demonstrates that ioGlutamatergic Neurons APP KM670/671NL hom and the genetically matched control (WT Control) lack the expression of pluripotency markers (NANOG and OCT4) at day 11, whilst robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic markers (VGLUT1 and VGLUT2), 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 / KM670/671NL following deterministic programming
bit.bio APP gene expression in ipsc derived glutamatergic neurons Alzheimer's disease in vitro cell model by RT-qPCR
Gene expression analysis demonstrates that ioGlutamatergic Neurons APP KM670/671NL hom and the genetically matched control (WT Control) express the APP gene encoding the amyloid precursor protein. Gene expression levels were assessed by RT-qPCR (data normalised to HMBS). Data represents day 11 post-revival samples, n=2 replicates.

Cells arrive ready to plate

bit.bio Glutamatergic neuron culture protocol is quick and uses open source media, cells mature rapidly

ioGlutamatergic Neurons APP KM670/671NL hom 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

bit.bio Carry out glutamatergic neuron culture in 96-well and 384-well plates for screening applications

The recommended minimum seeding density is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other similar commercially available products. 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, 48, 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

Homozygous KM670/671NL double point mutation (Swedish) in the APP gene

Applications

Alzheimer's disease research
Drug discovery and development
Disease modelling

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
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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
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ioGlutamatergic Neurons Brochure
ioGlutamatergic Neurons

bit.bio

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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

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Circadian clocks in human cerebral organoids Publication
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Rzechorzek, et al

bioRxiv

2024

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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
Smith, et al. 
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2024
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Schizophrenia risk gene ZNF804A controls ribosome localization and synaptogenesis in developing human neurons Publication
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2024

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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

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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
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2023

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Industrialising Cellular Reprogramming: Leveraging opti-ox Technology to Manufacture Human Cells with Unprecedented Consistency Talk
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HNRNPH1 regulates the neuroprotective cold‐shock protein RBM3 expression through poison exon exclusion Publication
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Cell culture hacks | human iPSC-derived glutamatergic neurons

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

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