Product nameioNEURONS/glut – Human iPSC-Derived Glutamatergic Neurons
DescriptionioNEURONS/glut - Human iPSC-Derived Glutamatergic Neurons
Tested applicationsSuitable for: High throughput screening, Functional Studies, ICC/IF, CellDiffmore details
For non-profit organizations, please use discount code ACCESS-TMA72 for a discounted price.
Product is available in three sizes:
Small - 7.5x105 cells/vial
Large - 1.5x106 cells/vial
Extra large - 6 x106 cells (4 x Large vials, at 1.5x106 cells/vial)
ioNEURONS/glut cells have been reprogrammed from human induced pluripotent stem cells (iPSC) using opti-ox, a precise reprogramming technology. Human stem cells, within days, convert into consistent, mature, functional glutamatergic neurons providing a high quality human model for the study of neurological activity and disease.
ioNEURONS/glut cultures consist mainly of glutamatergic neurons (>80%) characterised by the expression of the glutamate transporter genes VGLUT1 and VGLUT2 (see Figure 1). The minor remaining fraction of the neuronal population express marker genes of cholinergic neurons. A bulk RNA-seq analysis shows that ioNEURONS/glut have a rostral CNS identity and express the classical cortical maker genes FOXG1 and TBR1 (data not shown).
In partnership with Bit Bio Ltd.
Starting material: Human iPSC line
Seeding Density: 30,000 cells/cm2
Seeding compatibility: 24-, 96- and 384-well compatible
Quality control: ICC and gene expression analysis
Product applications: Academic research, Drug development, Neurotoxicology, High-throughput screening, Genetic screening (e.g. CRISPR screening).
This product is subject to limited use licenses from iPS Academia Japan Inc, TET Systems GmbH, ERS Genomics Limited and Sigma-Aldrich Co. LLC and is developed with Bit Bio patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Number of cells1.5 x 106 cells/vial
Storage instructionsShipped on Dry Ice. Store in liquid nitrogen.
Storage bufferConstituent: 10% DMSO
- CNQX, AMPA / kainate antagonist (ab120017)
- Tetrodotoxin, Na+ channel blocker (ab120054)
- Y-27632 dihydrochloride, Rho kinase inhibitor (ab120129)
- Doxycycline hyclate, matrix metalloprotease inhibitor (ab141091)
- Cytarabine, Pyrimidine nucleoside analog (ab141924)
- Anti-VGLUT2 antibody [EPR21085] (ab216463)
- Anti-VGluT1 antibody [EPR22269] (ab227805)
Our Abpromise guarantee covers the use of ab259259 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|High throughput screening||Use at an assay dependent concentration.|
|Functional Studies||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|CellDiff||Use at an assay dependent concentration.|
Cost effective and flexible.
ioNEURONS/glut 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.
- Recommended seeding density for ioNEURONS/glut is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other available products on the market.
- One Small vial (0.75 x 106 viable cells) can plate a minimum of 0.5 x 24-well plate, 0.75 x 96-well plate, or 1 x 384-well plate.
- One Large vial (1.5 x 106 viable cells) you can plate a minimum of 1 x 24-well plate, 1.5 x 96-well plate, or 2 x 384-well plates.
Immunocytochemistry of ioNEURONS/glut at day 9 post plating shows homogeneous expression of pan neuronal marker MAP2 and a dense neuronal network. Image courtesy of Charles River Laboratories.
Immunofluorescent staining on post-revival day 11 demonstrates homogenous expression of pan-neuronal proteins (MAP2 and TUBB3) and glutamatergic neuron-specific transporters (VGLUT1 and VGLUT2). Cells exhibit neurite outgrowth.
ioNEURONS/glut after revival over the course of the first 11 days of culture. Day 1 to 11 post-thawing; 400X magnification; scale bar: 100µm
ioNEURONS/glut display neuronal activity that matures over-time. Examples of MaxOne high-resolution multi electrode array (MEA) recordings of ioNEURONS/glut in BrainPhys™ media. The activity maps show firing rate (A), spike amplitude (B) and % of active electrodes (C). Results demonstrate a time-dependent increase of spontaneous activity during neuronal maturation from 2 to 3 weeks post-revival. Iovino et al., Molecular and functional characterization of stem cells-derived glutamatergic neurons ioNEURONS/glut in support of drug discovery applications., Charles River Laboratories.
ioNEURONS/glut show good suitability for high-throughput screening in 384-well format plates. Cytotoxicity CellTiter-Glo®️ (CTG) and TR-FRET (HTRF®️) assays for AKT serine/threonine kinase 1 (AKT) and Huntingtin (HTT) proteins were performed on ioNEURONS/glut in 384-well plates treated with tool compound (cmp) at day 9 post-revival. Compound titration results in a concentration response curve for all three assays (mean±sd of 2 replicates). CTG assay on ioNEURONS/glut shows an excellent average signal/ background ratio and high suitability for HTS. HTRF®assays on ioNEURONS/glut show lower signals but with low variability, and could therefore also provide a suitable platform for HTS. Iovino et al., Molecular and functional characterization of stem cells-derived glutamatergic neurons ioNEURONS/glut in support of drug discovery applications., Charles River Laboratories.
ioNEURONS/glut 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: 1. Induction (carried out at Bit Bio) 2. Stabilization for 4 days with Doxycycline 3. Maintenance during which the neurons mature.
ab259259 has not yet been referenced specifically in any publications.