樱桃视频

ioGlutamatergic-neurons-MAPT-N279K-hom-DAPI-MAP2-VGLUT2-overlay-1

cat no | io1014

ioGlutamatergic Neurons
MAPT N279K/N279K

Human iPSC-derived FTD disease model

A rapidly maturing, consistent and scalable system to study frontotemporal dementia (FTD).

ioGlutamatergic Neurons MAPT N279K/N279K are opti-ox deterministicly programmed glutamatergic neurons carrying a genetically engineered homozygous N279K mutation in the MAPT gene encoding the microtubule-associated protein tau.

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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.

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Benchtop benefits

phenotype_0

Disease related phenotype

High content ICC image analysis shows hyperphosphorylation of tau compared to the wild-type control.

comparison_0

Make True Comparisons

Pair the ioDisease Model Cells with the genetically matched wild-type ioGlutamatergic Neurons to directly investigate the impact of mutant tau protein on disease.

quick_0

Quick

The disease model cells and genetically matched control are experiment ready as early as 2 days post revival, and form structural neuronal networks at 11 days.

Technical data
Technical data
Product information Supporting documentation Resources Related products

Disease-related phenotype

Hyperphosphorylation of tau observed in the disease model cells compared to the wild type control

Hyperphosphorylation of tau in glutamatergic neurons FTD disease model cells carrying MAPT P301S or N279K mutation

ioGlutamatergic Neurons disease model cells carrying MAPT P301S/P301S, MAPT N279K/WT and MAPT N279K/N279K mutations show hyperphosphorylation when compared to the wild type control ioGlutamatergic Neurons (WT) at day 21. The bar graphs show total Tau, pTau217/total Tau, pTau202/5/total Tau or pTau404/total Tau in cell bodies, analysed by immunocytochemistry. Statistical analyses performed on 5 cellular replicates in the same plate. Bars showing mean, error bars showing standard deviation. Statistics calculated by one way ANOVA and Tukey posthoc analysis. Data courtesy of Charles River Laboratories.

Highly characterised and defined

ioGlutamatergic Neurons MAPT N279K/N279K express neuron-specific markers comparably to the wild type control

ioGlutamatergic Neurons MAPT N279K/N279K disease model cells express neuron-specific markers comparably to 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 MAPT N279K/N279K compared to the genetically matched control. 100X magnification.

ioGlutamatergic Neurons MAPT N279K/N279K form structural neuronal networks by day 11

ioGlutamatergic Neurons MAPT N279K/N279K disease model cells morphology by brightfield imaging over 11 days post-revival

ioGlutamatergic Neurons MAPT N279K/N279K mature rapidly 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 MAPT N279K/N279K demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following v programming

ioGlutamatergic Neurons MAPT N279K/N279K key neuronal marker gene expression analysis by RT-qPCR

Gene expression analysis demonstrates that ioGlutamatergic Neurons MAPT N279K/N279K 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 is shown relative to the parental human iPSC control (hiPSC), normalised to HMBS. Data represents day 11 post-revival samples; n=2 biological replicates.

Disease-related MAPT is expressed in ioGlutamatergic Neurons MAPT N279K/N279K following deterministic programming

ioGlutamatergic Neurons MAPT N279K/N279K expression analysis of MAPT gene by RT-qPCR

RT-qPCR analysis demonstrates a similar expression level of the MAPT gene in both wild type ioGlutamatergic Neurons (WT Control) and ioGlutamatergic Neurons MAPT N279K/N279K at day 11 post-revival (n=2 replicates). cDNA samples of the parental human iPSC line (hiPSC) were included as a reference.

Cells arrive ready to plate

ioGlutamatergic Neurons MAPT N279K/N279K arrive ready to plate

ioGlutamatergic Neurons MAPT N279K/N279K 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

Do more with every vial

ioGlutamatergic_Neurons_seeding_density_small_96_384

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, 48, 96 & 384 well plates

Shipping info

Dry ice

Donor

Caucasian adult male (skin fibroblast)

Vial size

Small: >1 x 10鈦� 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掳C

Format

Cryopreserved cells

Genetic modification

Homozygous N279K missense mutation in the MAPT gene

Applications

FTD research
Drug discovery and development
Disease modelling
High content imaging
Western blotting
Electrophysiological assays (MEA)
Co-culture studies

Product use

ioCells are for research use only

*ioGlutamatergic Neurons MAPT N279K/N279K disease model cells can be used with the parental wild type ioGlutamatergic Neurons (io1001) as a control. The two products are genetically matched except for a ~850Kb amplification on 20q11.21 in the disease model cells, identified by Optical Genome Mapping.

Product resources

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Human iPSC-Based Models of Glial Cells for Studying Neurodegenerative Disease Webinar
Human iPSC-Based Models of Glial Cells for Studying Neurodegenerative Disease
Valentina Fossati, PhD | Senior Research Investigator | The New York Stem Cell Foundation

In锚s Ferreira | Senior Product Manager | 樱桃视频
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ioGlutamatergic Neurons Brochure
ioGlutamatergic Neurons

樱桃视频

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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

樱桃视频

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鈥檚 disease Poster
Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer鈥檚 disease
Smith, et al. 
樱桃视频
2024
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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鈥檚 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 | 樱桃视频

Javier Conde-Vancells | Director Product Management | 樱桃视频

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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 | 樱桃视频

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Cell culture hacks for human iPSC-derived glutamatergic neurons

Read this blog for top tips on cell culture of human iPSC-derived glutamatergic neurons to help you overcome some of the most common issues. By using careful handling, it鈥檚 possible to achieve regular and consistent cell culture success!

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Cell culture hacks for ioGlutamatergic Neurons

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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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