bit.bio-ioGlutamatergic-Neurons

cat no | io1001

ioGlutamatergic Neurons

Human iPSC-derived glutamatergic neurons

ioGlutamatergic Neurons from human induced pluripotent stem cells (iPSC) deterministically programmed using opti-ox technology. Within days, cells convert consistently to mature, functional glutamatergic neurons characterised by  >80% expression of glutamate transporter genes VGLUT1 and VGLUT2.

Place your order

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.

per vial

Benchtop benefits

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Quick

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

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Scalable

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

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Easy to use

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

Technical data

Ready within days

ioGlutamatergic Neurons generated by transcription factor-driven deterministic programming of iPSCs using opti-ox technology

Time-lapse video capturing the rapid and homogeneous neuronal phenotype acquisition upon thawing of cryopreserved ioGlutamatergic Neurons. 7 day time course.

Highly characterised and defined

ioGlutamatergic Neurons express glutamatergic neuron-specific markers

bit.bio ioGlutamatergic Neurons - Day 11 - MAP2 square
MAP2
bit.bio ioGlutamatergic Neurons - Day 11 - VGLUT2square
VGLUT2
bit.bio ioGlutamatergic Neurons - Day 11 - DAPIsquare
DAPI
bit-bio ioGlutamatergic Neurons - Day 11 - MERGE VGLUT2(g) MAP2(r) DAPI(b)
MERGE

Immunofluorescent staining on post-revival day 11 demonstrates homogenous expression of the pan-neuronal protein, MAP2 and glutamatergic neuron-specific transporter, VGLUT2. 

ioGlutamatergic Neurons form structural neuronal networks by day 11

WT isogenic control brightfield day 1
Day 1
WT isogenic control brightfield day 4
Day 4
WT isogenic control brightfield Day 7_1
Day 7
WT isogenic control brightfield day 11
Day 11

ioGlutamatergic Neurons mature rapidly and form structural neuronal networks over 11 days. Day 1 to 11 post thawing; 100X magnification.

Whole transcriptome analysis demonstrates high lot-to-lot consistency across three manufactured lots of ioGlutamatergic Neurons

Bulk RNA-seq data represented in PCA plots demonstrates lot-to-lot consistency of 3 different lots of ioGlutamatergic Neurons.

Bulk RNA-sequencing analysis was performed on three different lots of ioGlutamatergic Neurons on day 0, day 11 and day 18 post-revival. (A) A principal component analysis (PCA) to assess gene expression variance between three different manufactured lots showed a tight clustering of the samples at each timepoint, demonstrating high consistency between these lots. This lot-to-lot consistency of ioGlutamatergic Neurons will help reduce experimental variation and increase the reproducibility of experiments. (B) PCA without the parental non-induced hiPSC samples, highlighting the tight clustering of the day 11 as well as day 18 samples of the three different lots. (C) Differential expression test reveals no statistically significant differentially expressed (DE) genes across the three lots at day 11 (|logFC| > 0.5 and FDR < 0.01).

Colours represent the three lots of products; shapes represent the parental non-induced hiPSC line and different timepoints.

Expression levels for specific genes of interest can be requested by contacting our team at technical@bit.bio.

High lot-to-lot consistency is demonstrated by a consistent transcriptomic fingerprint across manufactured lots of ioGlutamatergic Neurons

Single cell RNA-seq data represented in UMAP plots demonstrates lot-to-lot consistency of 3 different lots of ioGlutamatergic Neurons.
Single cell RNA-sequencing analysis was performed on three different lots of ioGlutamatergic Neurons on day 11. UMAP plots represent the cell-to-cell variation in gene expression profiles of cells, each dot representing an individual cell. Cells from each of the three lots are equally distributed across the body of the plot. Merging the UMAP plots creates a tight overlay, showing a strong transcriptional relationship between cells from three independently manufactured lots of ioGlutamatergic Neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing.

Single cell ATAC-sequencing shows a consistent transcriptomic fingerprint demonstrating high lot-to-lot consistency across manufactured lots of ioGlutamatergic Neurons

Single cell ATAC-seq data represented in UMAP plots demonstrates lot-to-lot consistency of 3 different lots of ioGlutamatergic Neurons.
Single cell ATAC-sequencing analysis was performed on three different lots of ioGlutamatergic Neurons on day 11. Single cell ATAC-sequencing reveals regions of open chromatin to understand the gene regulatory landscape of individual cells. UMAP plots represent the cell-to-cell variation in chromatin accessibility of the cells, each dot representing a single cell. Cells from each of the three lots are equally distributed across the body of the plot. Merging the UMAP plots creates a tight overlay, showing a strong transcriptional relationship between cells from three independently manufactured lots of ioGlutamatergic Neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing.

Rapid gain of functional activity

ioGlutamatergic Neurons display neuronal activity that matures over-time

ioGlut-MEA

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

ioGlut-spontaneous-burst-A
ioGlut-spontaneous-burst-B
ioGlut-spontaneous-burst-C

Click on the tabs to explore the data.

(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

ioGlut-AWSDR-D8
ioGlut-AWSDR-D13
ioGlut-AWSDR-D20

Click on the tabs to explore the data.

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.

Robust and scalable cells for high-throughput screening

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

ioGlut-HTS

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.

 

Industry leading seeding density

Do more with every vial

ioGlut-WT-well_plate-2

The recommended minimum seeding density is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other similar 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 plates. One large vial can plate a minimum of 3.6 x 24-well plates, 5.4 x 96-well plates, or 7.75 x 384-well plates. This means every vial goes further, enabling more experimental conditions and more repeats, resulting in more confidence in the data.

Easy culturing

Cells arrive ready to plate

bit.bio_ioGlutamatergic_Neurons_WT_timeline

ioGlutamatergic Neurons 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: 1. Stabilisation for 4 days 2. Maintenance during which the neurons mature.

Product information

Starting material

Human iPSC line

Karyotype

Normal (46, XY)

Seeding compatibility

6, 12, 24, 48, 96 & 384 well plates

Shipping info

Dry ice

Donor

Caucasian adult male (skin fibroblast)

Vial size

Small: >1 x 10 viable cells Large: >5 x 10 viable cells

Quality control

Sterility, protein expression (ICC) and gene expression (RT-qPCR)

Differentiation method

opti-ox deterministic cell programming

Recommended seeding density

30,000 cells/cm²

User storage

LN2 or -150°C

Format

Cryopreserved cells

Product use

ioCells are for research use only

Applications

Drug discovery
Neurotoxicology
High throughput screening
CRISPR Screening
3D bioprinting

Product resources

ioGlutamatergic Neurons Brochure
ioGlutamatergic Neurons

bit.bio

Download
ioGlutamatergic Neurons Wild Type and related disease models | User Manual User manual
ioGlutamatergic Neurons Wild Type and related disease models | User Manual

V11

bit.bio

2024

Download
Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease Poster
Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease
Smith et al. 
bit.bio
2024
Download
Generating publishable neuroscience research in 12 weeks with ioGlutamatergic Neurons Case study
Generating publishable neuroscience research in 12 weeks with ioGlutamatergic Neurons

Professor Deepak Srivastava

Professor of Molecular Neuroscience and Group Leader, MRC Centre for Developmental Disorders

King’s College London 

Download
Running Large-Scale CRISPR Screens in Human Neurons Webinar
Running Large-Scale CRISPR Screens in Human Neurons

Emmanouil Metzakopian | Vice President, Research and Development | bit.bio

Javier Conde-Vancells | Director Product Management | bit.bio

Watch now
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming Webinar
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming

Dr Ania Wilczynska | Head of Computational Genomics | Non-Clinical | bit.bio

Watch now
Industrialising Cellular Reprogramming: Leveraging opti-ox Technology to Manufacture Human Cells with Unprecedented Consistency Talk
Industrialising Cellular Reprogramming: Leveraging opti-ox Technology to Manufacture Human Cells with Unprecedented Consistency

Innovation showcase talk at ISSCR

Marius Wernig MD, PhD | Stanford 

Mark Kotter, MD, PhD | bit.bio

Watch now
Modelling neurodegeneration: Human isogenic system to study FTD & ALS Poster
Modelling neurodegeneration: Human isogenic system to study FTD & ALS

Oosterveen, et al

bit.bio & Charles River Laboratories

2023

View

Cell culture hacks | human iPSC-derived glutamatergic neurons

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.

bit.bio_3x2_ioGlutamatergic Neurons_MAP2_Hoescht_x20_hi.res (1)

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Wild type and isogenic disease model cells: A true comparison

Be confident in your data by pairing ioDisease Model cells with the genetically matched ioWild Type control

 

bitbio-vials-Wild_and_Disease-staggered-2500px_wide-B

Related pages

Discover ioCells Learn about our range of human iPSC-derived cells for research and drug discovery
Resources Explore our latest scientific insights, webinars, blogs and videos
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