A rapidly maturing, consistent and scalable system to study frontotemporal dementia (FTD).
ioGlutamatergic Neurons MAPT N279K/N279K are opti-ox™ precision reprogrammed glutamatergic neurons carrying a genetically engineered homozygous N279K mutation in the MAPT gene encoding the microtubule-associated protein tau.
The disease model cells show a FTD disease-related phenotype, indicated by hyperphosphorylation of tau compared to the genetically matched control.
Hyperphosphorylation of tau observed in the disease model cells compared to the wild type control
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.
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 genetically matched 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 N279K/N279K express neuron-specific markers comparably to the 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 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 reprogramming
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 reprogramming
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 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 MAPT N279K/N279K 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
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 N279K 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
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.
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’s possible to achieve regular and consistent cell culture success!