ioGlutamatergic Neurons

Human iPSC-derived glutamatergic neurons

io1001  |  Formerly known as ioNEURONS/glut, cat no: e001

ioGlutamatergic Neurons,  part of our ioCells portfolio, have been reprogrammed from human induced pluripotent stem cells (iPSC) using our precise reprogramming technology: opti-ox™¹ (optimized inducible overexpression). 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.

ioGlutamatergic Neurons consist mainly of glutamatergic neurons (>80%) characterised by the expression of the glutamate transporter genes VGLUT1 and VGLUT2. The minor remaining fraction of the neuronal population express marker genes of cholinergic neurons. A bulk RNA-seq analysis shows that ioGlutamatergic Neurons have a rostral CNS identity and express the classical cortical marker genes FOXG1 and TBR1 (data not shown).

Ready-to-culture cells are suitable as models for research in cell-type specific biology, target validation and drug screening in pharmaceutical R&D, and toxicology testing.

¹ Pawlowski et al., Stem Cell Report 2017

Product specifications


Ready for experimentation within days
Highly characterised and defined
Early electrical signal
Easy culturing


Academic research
– Drug development
– Co-culture studies
– High-throughput screening
Single cell CRISPR Screening
3D bioprinting

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Batch to batch reproducibility and homogeneity create a stable human model for excitatory neuronal activity and disease.


Ready for experimentation as early as 2 days post revival and form functional neuronal networks at 17 days.


Industrial scale quantities at a price point that allows the cells to be used from research to screening scale.


Cells arrive programmed to rapidly mature upon revival. One medium required in a two-step protocol.

ioGlutamatergic Neurons generated by NGN2-driven reprogramming of iPSCs using opti-ox™ technology

Video capturing the rapid morphological changes from iPSCs upon induction of NGN2 expression using opti-ox™ cellular reprogramming. 7 day time course.

ioGlutamatergic Neurons express glutamatergic neuron-specific markers

ioCells TUBB3
ioCells VGLUT1
ioCells MAP2
ioCells VGLUT2
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.

ioGlutamatergic Neurons after revival over the course of the first 11 days

ioCells ioGlutamatergic Neurons Day 1
ioCells ioGlutamatergic Neurons Day 4
ioCells ioGlutamatergic Neurons Day 7
ioCells ioGlutamatergic Neurons Day 11
Day 1 to 11 post-thawing; 400X magnification; scale bar: 100µm.

ioGlutamatergic Neurons display neuronal activity that matures over-time

ioCells ioGluneuronal activity

Examples of MaxOne high-resolution multi electrode array (MEA) recordings of ioGlutamatergic Neurons 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, M. et al., 2019, Charles River Laboratories.

Cells demonstrate a time-dependent increase of spontaneous bursting activity over a three-week period

A Percentage of active electrodes bursting

B Example of a spontaneous spike

C Example of a bursting phenotype

(A) The graph shows the % of active bursting electrodes for each time point. (B) An example of a spontaneous spike, taken at Day 8 post-revival (1 second sweep, 32 µV/-18 µV). (C) An example of a bursting phenotype, taken at Day 20 post-revival (1 second sweep, 16 µV/-16 µV). Cells were cultured in the recommended open-source medium and recorded on 64-electrode MEAs.

NDimension (Science and Engineering) Ltd.

ioGlutamatergic Neurons co-cultured with rat-derived astrocytes demonstrate time-dependent increase in synchronous activity

neurons MEA Synchronicity Day 8

neurons MEA Synchronicity Day 13

neurons MEA Synchronicity Day 20

Array Wide Spike Detection Rate histograms (AWSDR – a graphical measure of synchrony) for 10-minute recordings on Day 8, 13 and 20 post-revival ioGlutamatergic Neurons in co-culture with primary rat-derived astrocytes. Results show prominent synchronicity on Day 13, exemplified by the ‘spikier’ nature of the associated AWSDR, which increases at Day 20. Cells were cultured in the recommended open-source medium and recorded on 64-electrode MEAs.

NDimension (Science and Engineering) Ltd.

ioGlutamatergic Neurons show good suitability for high-throughput screening in 384-well format plates

ioCells ioGlutamatergic Neurons

Cytotoxicity CellTiter-Glo®️ (CTG) and TR-FRET (HTRF®️) assays for AKT serine/threonine kinase 1 (AKT) and Huntingtin (HTT) proteins were performed on ioGlutamatergic Neurons 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 ioGlutamatergic Neurons shows an excellent average signal/ background ratio and high suitability for HTS. HTRF® assays on ioGlutamatergic Neurons show lower signals but with low variability, and could therefore also provide a suitable platform for HTS.

Iovino, M. et al., 2019, Charles River Laboratories.

 Cells arrive ready to plate
bit bio ioGlutamatergic Neurons vial of cells

ioGlutamatergic Neurons 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: Phase 0. Induction (carried out at Phase 1. Stabilisation for 4 days. Phase 2. Maintenance during which the neurons mature.

Cost effective and flexible

bitbio-well_plate_diagram?i oGlut_Neurons–landscape
ioGlutamatergic Neurons 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 ioGlutamatergic Neurons is 30,000 cells/cm2, compared to up to 250,000 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.

Product specifications

Starting material
Human iPSC line

Caucasian adult male
(skin fibroblast)

Differentiation method
opti-ox™ cellular reprogramming

Normal (46, XY)

Vial size
Small: >1 x 106 viable cells
Large: >5 x 106 viable cells

Recommended seeding density
30,000 cells/cm2

Seeding compatibility
6 to 384 well plates

Quality control
Sterility, protein expression (IF) and gene expression (RT-qPCR)

User storage
LN2 or -150°C

Shipping info
Dry ice

Product use
These cells are for research use only

Supporting documentation