Batch to batch reproducibility and homogeneity create a stable human model for excitatory neuronal activity and disease.
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.
Advantages
– Ready for experimentation within days
– Highly characterised and defined
– Early electrical signal
– Easy culturing
Applications
– Academic research
– Drug development
– Co-culture studies
– High-throughput screening
– Single cell CRISPR Screening
– 3D bioprinting
For more information download our flyer
or email info@bit.bio
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 express glutamatergic neuron-specific markers
ioGlutamatergic Neurons after revival over the course of the first 11 days
ioGlutamatergic Neurons display neuronal activity that matures over-time
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) 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 bit.bio 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
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 bit.bio 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
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
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 bit.bio) Phase 1. Stabilisation for 4 days. Phase 2. Maintenance during which the neurons mature.
“Our major surprise when we first used the ioGlutamatergic Neurons was that after thawing the cells in 384-well format, we could see immediately after 2 days a nice neuronal network, and there was no well to well variability within the same plate. This made our assay quite robust.”
Dr Mariangela Iovino
Group Leader | Biology Discovery | Charles River
"These cells enable us to move rapidly as from the moment of plating within 4-7 days we have mature and functional neurons."
Dr Shushant Jain
Group Leader | In Vitro Biology | Charles River
"These cells provide a reliable and pure source of glutamatergic neurons, resembling primary human ones. They are ready-to-use which makes it much more easy for tissue culture work and for reproducible results.”
Dr Koby Baranes
Research Associate | University of Cambridge
"ioGlutamatergic Neurons are easy to use with a simple application protocol. They recover quickly after thaw and are able to form a mature mesh of neurons ideal for testing within a few days."
Dr Jeremy Anton
Scientist | Charles River
Starting material
Human iPSC line
Donor
Caucasian adult male
(skin fibroblast)
Differentiation method
opti-ox™ cellular reprogramming
Karyotype
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