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.
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 ofneuronal-specific and glutamatergic-specific markers following reprogramming
Gene expression analysis demonstrates that ioGlutamatergic Neurons TDP‑43 M337V/M337Vand the isogenic control (WT) lack the expression of pluripotency makers (NANOG and OCT4), at day 11, whilst robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic-specific (VGLUT1 and VGLUT2) markers, and the glutamate receptor GRIA4. Gene expression levels were assessed by RT-qPCR (data expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.
Disease-related TARDBP is expressed in ioGlutamatergic Neurons TDP‑43 M337V/M337V following reprogramming
Gene expression analysis demonstrates that ioGlutamatergic Neurons TDP‑43 M337V/M337Vand the isogenic control (WT) express the TARDBP gene encoding TDP‑43. Gene expression levels were assessed by RT-qPCR (data expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.
ioGlutamatergic Neurons TDP-43 M337V/M337V show reduced neuronal activity indicating their potential as a relevant translational in vitro drug discovery model for ALS and FTD
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.
Cells arrive ready to plate
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.
Industry leading seeding density
Do more with every vial
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.
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), genotype (Sanger seq) and gene expression (RT-qPCR)
These cells are for research use only
opti-ox cellular reprogramming
Recommended seeding density
LN2 or -150°C
FTD and ALS research Drug discovery and development Disease modelling High-throughput screening Electrophysiological assays (MEA) Co-culture studies
Homozygous M337V missense mutation in the TARDBP gene