cat no | io1007
Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
Make True Comparisons
Pair the ioDisease Model Cells with the genetically matched wild-type ioGlutamatergic Neurons to investigate the impact of the GBA mutation on molecular mechanisms and cell function.
Scalable
With opti-ox technology, we can make billions of consistently programmed cells, surpassing the demands of industrial workflows.
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 GBA null/R159W 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 GBA null/R159W express neuron-specific markers comparably to the isogenic control
Immunofluorescent staining on post-revival day 11 demonstrates similar homogenous expression of pan-neuronal proteins TUBB3 and MAP2 (upper panel) and glutamatergic neuron-specific transporter VGLUT2 (lower panel) in ioGlutamatergic Neurons GBA null/R159W compared to the isogenic control. 100X magnification.
ioGlutamatergic Neurons GBA null/R159W form structural neuronal networks by day 11
ioGlutamatergic Neurons GBA null/R159W 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 GBA null/R159W demonstrate gene expression of neuronal and glutamatergic-specific markers following deterministic programming
Gene expression analysis demonstrates that ioGlutamatergic Neurons GBA null/R159W and the isogenic control (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 normalised to HMBS; cDNA samples of the parental human iPSC line (hiPSC) were included as reference). Data represents day 11 post-revival samples, n=2 replicates.
Disease-related GBA is expressed in ioGlutamatergic Neurons GBA null/R159W following deterministic programming
Gene expression analysis demonstrates that ioGlutamatergic Neurons GBA null/R159W and the isogenic control (WT Control) express the GBA gene encoding the glucocerebrosidase protein. Gene expression levels were assessed by RT-qPCR (data normalised to HMBS, cDNA samples of the parental human iPSC line (hiPSC) were included as reference). Data represents day 11 post-revival samples, n=2 replicates.
GBA protein is present in ioGlutamatergic Neurons GBA null/R159W at a lower level than the wild type control
A Western blot experiment confirmed the presence of the GBA protein in ioGlutamatergic Neurons GBA null/R159W at a lower level than in the wild-type ioGlutamatergic Neurons. Day 11 cell lysates were subjected to Western blotting (20 µg protein in 40 µl per lane) using 4-20% mini protean TGX stain-free gels. Proteins were transferred onto PVDF membranes using the Trans-Blot Turbo Transfer Pack, blocked for 10 minutes, incubated with primary antibodies (GBA Invitrogen MA5-26589, 1:2000; GAPDH Abcam ab8245, 1:5000), washed three times, incubated with HRP-labelled secondary antibodies, washed three times and signal visualised by electrochemiluminescence.
1= ioGlutamatergic Neurons (wild type), 2= ioGlutamatergic Neurons GBA null/R159W.
The recommended minimum seeding density is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other similar commercially available products. 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. This means every vial goes further, enabling more experimental conditions and more repeats, resulting in more confidence in the data.
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Read this blog on glutamatergic neuron cell culture for our top tips on careful handling, cell plating and media changes to achieve success from the outset.
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Further your disease research by pairing our wild type cells with isogenic disease models.
Consistent. Defined. Scalable.
