A rapidly maturing, consistent and scalable isogenic system to study frontotemporal dementia (FTD).
ioGlutamatergic Neurons MAPT P301S/P301S are opti-ox™ precision reprogrammed glutamatergic neurons containing a genetically engineered homozygous P301S mutation in the MAPT gene encoding the tau protein.
The disease model cells show a FTD disease-related phenotype, indicated by hyperphosphorylation of tau compared to the isogenic control.
Hyperphosphorylation of tau observed in the disease model cells compared to the isogenic control
ioGlutamatergic Neurons disease model cells carrying MAPT P301S/P301S, MAPT N279K/WT and MAPT N279K/N279K mutations show hyperphosphorylation when compared to the isogenic 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.
The expression of the tau 4R isoform transcript is relevant for the study of frontotemporal dementia (FTD). Bulk RNA-sequencing data for ioGlutamatergic Neurons, the isogenic control for the MAPT disease model cells, show an equimolar tau 3R/4R ratio at day 11, similar to the ratio in the adult human brain, making the wild type and disease model cells a suitable system for investigating FTD.
Highly characterised and defined
ioGlutamatergic Neurons MAPT P301S/P301S express neuron-specific markers comparably to the isogenic 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 P301S/P301S compared to the isogenic control. 100X magnification.
ioGlutamatergic Neurons MAPT P301S/P301S form structural neuronal networks by day 11
ioGlutamatergic Neurons MAPT P301S/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 MAPT P301S/P301Sdemonstrate gene expression of neuronal-specific and glutamatergic-specific markers following reprogramming
Gene expression analysis demonstrates that at day 11, ioGlutamatergic Neurons MAPT P301S/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 MAPT P301S/P301S following reprogramming
RT-qPCR analysis demonstrates similar expression level of the MAPT gene in both wild type ioGlutamatergic Neurons (WT) and ioGlutamatergic Neurons MAPT P301S/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 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 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.
Human iPSC line
Normal (46, XY)
6, 12, 24, 48, 96 & 384 well plates
Caucasian adult male (skin fibroblast)
Small: >1 x 10⁶ viable cells Large: >5 x 10⁶ viable cells
Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)
opti-ox cellular reprogramming
Recommended seeding density
LN2 or -150°C
Homozygous P301S missense mutation in the MAPT gene
FTD research Drug discovery and development Disease modelling High content imaging Western blotting Electrophysiological assays (MEA) Co-culture studies