cat no | io1001
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.
Quick
Experiment ready as early as 2 days post revival and show synchronised network activity by 31 days.
Scalable
Industrial scale quantities at a price point that allows the cells to be used from research to screening scale.
Easy to use
Cells arrive programmed to mature rapidly upon revival. One medium is required in a two-step protocol.
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.
Dr Shushant Jain
Group Leader | In Vitro Biology | Charles River, 2021
Dr Mariangela Iovino
Senior Group Leader | Biology Discovery | Charles River
Dr Koby Baranes
Research Associate | University of Cambridge
Dr Jeremy Anton
Scientist | Charles River
Professor Deepak Srivastava
Professor | Molecular Neuroscience | King’s College London and Group Leader | MRC Centre for Developmental Disorders
ioGlutamatergic Neurons generated by transcription factor-driven deterministic cell 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.
ioGlutamatergic Neurons express glutamatergic neuron-specific markers
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Immunofluorescent staining on day 11 post-revival demonstrates homogenous expression of the pan-neuronal protein, MAP2 and glutamatergic neuron-specific transporter, VGLUT2.
ioGlutamatergic Neurons form structural neuronal networks by day 11
ioGlutamatergic Neurons mature rapidly, show glutamatergic neuron morphology 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-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-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-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.
ioGlutamatergic Neurons display neuronal activity that matures over time
The function of ioGlutamatergic Neurons was investigated using the MaxTwo HD-MEA System.
The Axon Tracking Assay (left) shows examples of reconstructed axonal paths of travelling action potentials of individual iPSC-derived glutamatergic neurons. The assay reveals the spatial propagation of the neuronal action potential from the soma to distant axonal branches.
Total axon length (middle) and firing rate (right) increase over time, indicating that the cells are maturing. ioGlutamatergic Neurons were cultured with human iPSC-derived astrocytes.
Data courtesy of Charles River Laboratories and MaxWell Biosystems.
Rapid maturation of ioGlutamatergic Neurons leads to synchronised network activity by day 31
Raster plots generated using the MaxTwo HD-MEA System show the development of the neuronal network over time.
The plots show the dynamics of the network activity using 1,024 active electrodes. Each row represents an individual electrode and each blue dot indicates a spike detected at that electrode over a period of 300 seconds.
Spontaneous activity is observed at DIV 7. Clear synchronised bursting activity is observed by DIV 31, represented by blue vertical lines, followed by an overall drop in activity, seen as white lines.
Download our poster to see additional data that shows how ioGABAergic Neurons form functional neuronal networks with ioGlutamatergic Neurons in the presence of astrocytes, and how the tri-culture responds to bicuculline and diazepam.
ioGlutamatergic Neurons offer a rapidly maturing functional system that can be used to assess neuronal networks and the impact of a drug treatment or intervention.
Data courtesy of Charles River Laboratories and MaxWell Biosystems.
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-to-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.
Data courtesy of Charles River Laboratories.
ioGlutamatergic Neurons offer a robust, physiologically-relevant model for efficacy screening of candidate ASOs
Positive and negative control antisense oligonucleotides (ASOs) with gapmer chemistry were introduced into glutamatergic neurons by gymnosis. RT-qPCR was used to measure ASO-induced gene knockdown.
Data courtesy of Charles River Laboratories.
Do more with every vial
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.
ioGlutamatergic Neurons display neuronal activity that matures over time
The function of ioGlutamatergic Neurons was investigated using the MaxTwo HD-MEA System.
The Axon Tracking Assay (left) shows examples of reconstructed axonal paths of travelling action potentials of individual iPSC-derived glutamatergic neurons. The assay reveals the spatial propagation of the neuronal action potential from the soma to distant axonal branches.
Total axon length (middle) and firing rate (right) increase over time, indicating that the cells are maturing. ioGlutamatergic Neurons were cultured with human iPSC-derived astrocytes.
Data courtesy of Charles River Laboratories and MaxWell Biosystems.
Rapid maturation of ioGlutamatergic Neurons leads to synchronised network activity by day 31
Raster plots generated using the MaxTwo HD-MEA System show the development of the neuronal network over time.
The plots show the dynamics of the network activity using 1,024 active electrodes. Each row represents an individual electrode and each blue dot indicates a spike detected at that electrode over a period of 300 seconds.
Spontaneous activity is observed at DIV 7. Clear synchronised bursting activity is observed by DIV 31, represented by blue vertical lines, followed by an overall drop in activity, seen as white lines.
Download our poster to see additional data that shows how ioGABAergic Neurons form functional neuronal networks with ioGlutamatergic Neurons in the presence of astrocytes, and how the tri-culture responds to bicuculline and diazepam.
ioGlutamatergic Neurons offer a rapidly maturing functional system that can be used to assess neuronal networks and the impact of a drug treatment or intervention.
Data courtesy of Charles River Laboratories and MaxWell Biosystems.
ioGlutamatergic Neurons offer a robust, physiologically-relevant model for efficacy screening of candidate ASOs
Positive and negative control antisense oligonucleotides (ASOs) with gapmer chemistry were introduced into glutamatergic neurons by gymnosis. RT-qPCR was used to measure ASO-induced gene knockdown.
Data courtesy of Charles River Laboratories.
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-to-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.
Data courtesy of Charles River Laboratories.
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.
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Be confident in your data by pairing ioDisease Model cells with the genetically matched ioWild Type control
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