ioGlutamatergic Neurons MAPT P301S/P301S ICC

cat no | io1008

ioGlutamatergic Neurons

MAPT P301S/P301S

Human iPSC-derived FTD disease model

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

ioGlutamatergic Neurons MAPTP301S/P301S are opti-ox™ precision reprogrammed glutamatergic neurons containing a genetically engineered homozygous P301S mutation in the MAPT gene encoding the tau protein. 

Disease model cells with the MAPT mutation are also available with a heterozygous P301S mutation (cat no. io1015) and a heterozygous N279K mutation (cat no. io1009).

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Benchtop benefits

Make True Comparisons

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.

scalable_0

Scalable

Industrial scale quantities are available with industry-leading seeding densities, and at a price point that allows the cells to be used from research to high throughput screening.

Quick

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.

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 viable cells

Quality control

Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)

Product use

These cells are for research use only

Differentiation method

opti-ox cellular reprogramming

Recommended seeding density

30,000 cells/cm2

User storage

LN2 or -150°C

Format

Cryopreserved cells

Applications

FTD research
Drug discovery and development
Disease modelling
Electrophysiological assays (MEA)
Co-culture studies

Genetic modification

Homozygous P301S missense mutation in the MAPT gene

Technical data

Highly characterised and defined

ioGlutamatergic Neurons MAPTP301S/P301S express neuron-specific markers comparably to the isogenic control

ioGlutamatergic Neurons MAPT P301S/P301S ICC single channel and overlays

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 MAPTP301S/P301S compared to the isogenic control. 100X magnification.

ioGlutamatergic Neurons MAPTP301S/P301S form structural neuronal networks by day 11

ioGlutamatergic Neurons MAPT P301S/P301S Brightfield images

ioGlutamatergic Neurons MAPTP301S/P301S mature rapidly and form structural neuronal networks over 11 days, when compared to the isogenic control. Day 1 to 11 post thaw; 100X magnification.

ioGlutamatergic Neurons MAPTP301S/P301S demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following reprogramming

ioGlutamatergic Neurons MAPT P301S/P301S rt-qPCR of key markers

Gene expression analysis demonstrates that at day 11, ioGlutamatergic Neurons MAPTP301S/P301S (MAPT P301S/P301S) and the wild type isogenic control (WT) lack the expression of pluripotency makers (NANOG and OCT4) 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 hiPSC control (iPSC Control), normalised to HMBS. Data represents day 11 post-revival samples; n=2 biological replicates.

Disease-related MAPT is expressed in ioGlutamatergic Neurons MAPTP301S/P301S following reprogramming

ioGlutamatergic Neurons MAPT P301S/P301S rt-qPCR of MAPT

RT-qPCR analysis demonstrates similar expression level of the MAPT gene in both wild type ioGlutamatergic Neurons (WT) and ioGlutamatergic Neurons MAPTP301S/P301S (MAPT P301S/P301S) at day 11 post-revival (n=2 replicates). cDNA samples of the parental iPSC line (iPSC Control) were included as a reference.

Cells arrive ready to plate

ioGlutamatergic Neurons MAPT P301S/P301S arrive ready to plate

ioGlutamatergic Neurons MAPTP301S/P301S are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: Induction, which is carried out at bit.bio (Phase 0), Stabilisation for 4 days (Phase 1), and Maintenance (Phase 2) during which the neurons mature. Phases 1 and 2 after revival of cells are carried out at the customer site.

Industry leading seeding density

Do more with every vial
ioGlutamatergic Neurons MAPT P301S/P301S are compatible with 96 and 384 well plates

ioGlutamatergic Neurons MAPTP301S/P301S cells are compatible with plates ranging from 6 to 384 wells.

The recommended seeding density is 30,000 cells/cm2, compared to up to 500,000 cells/cm2 for other similar products on the market.

This means scientists are able to do more with every vial and expand experimental design within budget without losing out on quality. Resulting in more experimental conditions, more repeats, and more confidence in the data.

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 plate. 

Product resources

Modelling neurodevelopment | Investigating the impact of maternal immune activation on neurodevelopment using human stem cell models Webinar
Modelling neurodevelopment | Investigating the impact of maternal immune activation on neurodevelopment using human stem cell models

Dr Deepak Srivastava | King’s College London

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Partnering with Charles River to advance CNS drug discovery with ioGlutamatergic Neurons™

Charles River

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Dr Kaiser Karim | Scientist
bit.bio

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How to prepare tissue culture vessels with PDL-Geltrex™ for ioGlutamatergic Neuron™ culture

Dr Kaiser Karim | Scientist
bit.bio

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In Conversation with Charles River Video
In Conversation with Charles River

Dr Marijn Vlaming | Head of Biology
Charles River

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High Content Analysis of 2D and 3D cellular models for target and phenotypic drug discovery Poster
High Content Analysis of 2D and 3D cellular models for target and phenotypic drug discovery

Lachize | et al
Courtesy of Charles River Laboratories

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ioGlutamatergic Neurons™ Brochure
ioGlutamatergic Neurons™

bit.bio

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Read this blog to find out how experts from across academia and industry are approaching the challenges of reproducibility of in vitro cell models as well as potential solutions.

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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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