bit.bio-ioGlutamatergic-Neurons

cat no | io1001

ioGlutamatergic Neurons

Human iPSC-derived glutamatergic neurons

ioGlutamatergic Neurons have been precision reprogrammed from human induced pluripotent stem cells (iPSC) 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. 

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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 reprogramming 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.

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 seeding density of our human iPSC-derived glutamatergic neurons has been optimised and validated making it possible to have a cost point of under £0.67 (~$0.79) per well (96 well plate, seeding density 30,000 cells/cm2, Large vial size).

This means scientists are able to do more with every vial and expand experimental design within budget without losing out on quality. Resulting in more experimental conditions, more repeats, and more confidence in the data.

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.

Easy culturing

Cells arrive ready to plate

ioGlut-timeline

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: 1. Induction (carried out at bit.bio) 2. Stabilisation for 4 days 3. 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 cellular reprogramming

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

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
3D bioprinting of iPSC neuron-astrocyte coculture Publication
3D bioprinting of iPSC neuron-astrocyte coculture

Whitehouse et al.
JoVE Journal of Visualized Experiments 
2023

Using ioGlutamatergic Neurons

Read more
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

Dr Tony Oosterveen, et al.

bit.bio & Charles River Laboratories

2023

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

V7

bit.bio

2023

Download
Rethinking Developmental Biology With Cellular Reprogramming Webinar
Rethinking Developmental Biology With Cellular Reprogramming

Mark Kotter | CEO and founder | bit.bio

Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology |  Stanford University

Watch now
Precision Cellular Reprogramming for Scalable and Consistent Human Neurodegenerative Disease Models Talk
Precision Cellular Reprogramming for Scalable and Consistent Human Neurodegenerative Disease Models

Madeleine Garrett | Field Application Specialist | bit.bio

Watch now

Giving you easy access to endless and reliable human cells

Read this blog to find out how our precision cellular reprogramming technology, opti-ox is powering cell identity, giving you easy access to endless and reliable human cells!

Giving you easy access to the best cells

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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
Our platform Discover the cell identity coding platform behind our cells