cat no | ioEA1005
A rapidly maturing, consistent and scalable isogenic system to study amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD).
ioGlutamatergic Neurons TDP‑43 M337V/M337V are opti‑ox™ precision reprogrammed glutamatergic neurons carrying a genetically engineered homozygous M337V mutation in the TARDBP gene, encoding TAR DNA binding protein 43 (TDP‑43).
The disease model cells show a phenotype of reduced neuronal activity compared to the isogenic control detected by microelectrode array (MEA) analysis.
Related disease model cells are available with a heterozygous TDP‑43 M337V/WT mutation, and both can be used alongside their genetically matched (isogenic) control, ioGlutamatergic Neurons™.
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
Disease-related phenotype
MEA analysis detects lower weighted mean firing rate and network burst frequency compared to the wild‑type control.
Make True Comparisons
Pair the ioDisease Model Cells with the genetically matched wild-type ioGlutamatergic Neurons to investigate the impact of mutant TDP‑43 protein on disease progression.
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 TDP‑43 M337V/M337V express neuron-specific markers with protein expression highly reminiscent 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 TDP‑43 M337V/M337V compared to the isogenic control. 100X magnification.
ioGlutamatergic Neurons TDP‑43 M337V/M337V form structural neuronal networks by day 11
ioGlutamatergic Neurons TDP‑43 M337V/M337V mature rapidly and form structural neuronal networks over 11 days when compared to the isogenic control. Day 1 to 11 post-thawing; 100X magnification.
ioGlutamatergic Neurons TDP‑43 M337V/M337V demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following reprogramming
Reduced neuronal activity was measured in ioGlutamatergic Neurons TDP-43 M337V/M337V compared to ioGlutamatergic Neurons TDP-43 M337V/WT and the isogenic control, ioGlutamatergic Neurons. Microelectrode array (MEA) chips were spotted with 100K (~900K cells/cm2) ioGlutamatergic Neurons (WT), TDP-43 M337V/WT (C20), or TDP-43 M337V/M337V (C1), along with 20K (~180K cells/cm2) human iPSC-derived astrocytes. Brightfield at 26 DIV (A, left), cells show good coverage of electrodes and produce clear burst and network burst activity as seen in the raster plot of activity (A, right). In the raster plot, each dash indicates a firing event, blue indicates a single electrode burst and the pink box indicates a network burst event. Quantification of raster plots over the course of culture shows that ioGlutamatergic Neurons TDP-43 M337V/M337V have a lower weighted mean firing rate, and network burst frequency than WT and ioGlutamatergic Neurons TDP-43 M337V/WT (B). No clear difference is noted between WT and TDP-43 M337V/WT. Error bars indicate SEM, n=14 technical repeats. Data courtesy of Charles River Laboratories.
ioGlutamatergic Neurons TDP‑43 M337V/M337V 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 ioGlutamatergic Neurons TDP‑43 M337V/M337V mature. Phases 1 and 2 after revival of cells are carried out at the customer site.
ioGlutamatergic Neurons TDP‑43 M337V/M337V are compatible with plates ranging from 6 to 384 wells and are available in two vial sizes, tailored to suit your experimental needs with minimal waste.
The recommended seeding density is 30,000 cells/cm2, compared to up to 500,000 cells/cm2 for other available 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 plate. One Large vial can plate a minimum of 3.6 x 24-well plate, 5.4 x 96-well plate, 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), genotype (Sanger seq) and gene expression (RT-qPCR)
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 and ALS research
Drug discovery and development
Disease modelling
High-throughput screening
Electrophysiological assays (MEA)
Co-culture studies
Genetic modification
Homozygous M337V missense mutation in the TARDBP gene
Dr Tony Oosterveen, et al.
bit.bio & Charles River Laboratories
2023
Madeleine Garrett | Field Application Specialist | bit.bio
Oosterveen et al.
bit.bio
2022
Laila Ritsma et al.
Charles River Laboratories & bit.bio
2022
Dr Mariangela Iovino | Group Leader | Charles River
Dr Tony Oosterveen | Senior Scientist | bit.bio
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.