cat no | io1015
A rapidly maturing, consistent and scalable isogenic system to study frontotemporal dementia (FTD).
ioGlutamatergic Neurons MAPT P301S/WT are opti-ox™ precision reprogrammed glutamatergic neurons carrying a genetically engineered heterozygous P301S mutation in the MAPT gene encoding the tau protein.
These cells are part of a range that includes a homozygous MAPT P301S mutation, a heterozygous MAPT N279K mutation and a homozygous MAPT N279K mutation. When paired with the genetically matched (isogenic) control, wild type ioGlutamatergic Neurons™, these disease model cells offer a physiologically relevant model to investigate the impact of mutant tau protein on disease progression.
per vial
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@bit.bio.
Click here for bulk request
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
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
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.
ioGlutamatergic Neurons MAPT P301S/WT 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/WT compared to the isogenic control. 100X magnification.
ioGlutamatergic Neurons MAPT P301S/WT form structural neuronal networks by day 11
ioGlutamatergic Neurons MAPT P301S/WT 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/WT demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following reprogramming
Gene expression analysis demonstrates that at day 11, ioGlutamatergic Neurons MAPT P301S/WT (MAPT P301S/WT) and the 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/WT 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/WT (MAPT P301S/WT) at day 11 post-revival (n=2 replicates). cDNA samples of the parental iPSC line (iPSC Control) were included as a reference.
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 disease model cells carrying MAPT P301S/WT mutation do not show hyperphosphorylation. 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.
ioGlutamatergic Neurons MAPT P301S/WT 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.
ioGlutamatergic Neurons MAPT P301S/WT 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 plates. One Large vial can plate a minimum of 3.6 x 24-well plates, 5.4 x 96-well plates, or 7.75 x 384-well plates.
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 10⁶ viable cells
Large: >5 x 10⁶ viable cells
Quality control
Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)
Differentiation method
opti-ox cellular reprogramming
Recommended seeding density
30,000 cells/cm2
User storage
LN2 or -150°C
Format
Cryopreserved cells
Genetic modification
Heterozygous P301S 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
Emmanouil Metzakopian | Vice President, Research and Development | bit.bio
Javier Conde-Vancells | Director Product Management | bit.bio
Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | bit.bio
Innovation showcase talk at ISSCR
Marius Wernig MD, PhD | Stanford
Mark Kotter, MD, PhD | bit.bio
Dr Tony Oosterveen, et al.
bit.bio & Charles River Laboratories
2023
Mark Kotter | CEO and founder | bit.bio
Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology | Stanford University
Madeleine Garrett | Field Application Specialist | bit.bio
Laila Ritsma et al.
Charles River Laboratories & bit.bio
2022
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.
Further your disease research by pairing our wild type cells with isogenic disease models.