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bit.bio GFP glutamatergic neurons
Neuronal marker protein expression in GFP glutamatergic neurons
bit.bio-io1107-gfp-glutamatergic-neurons-GFP-expression-flow-cytometry
GFP Glutamatergic Neurons show expected neuronal morphology
bit.bio-io1107-gfp-glutamatergic-neurons-gene-expression-rt-qpcr
Co-culture workflow for GFP glutamatergic neurons and microglia
bit.bio GFP glutamatergic neurons
Neuronal marker protein expression in GFP glutamatergic neurons
bit.bio-io1107-gfp-glutamatergic-neurons-GFP-expression-flow-cytometry
GFP Glutamatergic Neurons show expected neuronal morphology
bit.bio-io1107-gfp-glutamatergic-neurons-gene-expression-rt-qpcr
Co-culture workflow for GFP glutamatergic neurons and microglia

cat no | io1107 Early Access

GFP ioGlutamatergic Neurons

Human iPSC-derived glutamatergic neurons constitutively expressing GFP

  • Glutamatergic neurons constitutively expressing GFP, easy to track in multi-cell cultures, ideal for live-cell imaging
  • Cryopreserved human iPSC-derived cells powered by opti-ox that are ready for experiments in days
  • Co-culture compatible with ioMicroglia and ioAstrocytes

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bit.bio GFP glutamatergic neurons

Human iPSC-derived glutamatergic neurons constitutively expressing GFP

Neuronal marker protein expression in GFP glutamatergic neurons

GFP ioGlutamatergic Neurons express key pan-neuronal and glutamatergic-specific markers

Immunofluorescent staining 11 days post-revival shows similar homogenous expression of pan-neuronal markers MAP2 and TUBB3 (upper panel) and the glutamatergic transporter VGLUT2 (lower panel) in both GFP ioGlutamatergic Neurons and the wild-type control.

The GFP signal is visible exclusively in GFP ioGlutamatergic Neurons and absent in the wild-type control (lower panel). 10X magnification.

bit.bio-io1107-gfp-glutamatergic-neurons-GFP-expression-flow-cytometry

Flow cytometry analysis of GFP expression in GFP ioGlutamatergic Neurons at day 11 and day 21

Flow cytometry analysis demonstrating GFP expression in >99% of cells for GFP ioGlutamatergic Neurons cultured until day 11 (centre), and no GFP expression seen in wild-type ioGlutamatergic Neurons (left).  At day 21, the percentage of cells expressing GFP has not decreased, indicating there is no silencing of the reporter gene. 

GFP Glutamatergic Neurons show expected neuronal morphology

GFP ioGlutamatergic Neurons form structural neuronal networks by day 11

GFP ioGlutamatergic Neurons mature rapidly, show glutamatergic neuron morphology and form structural neuronal networks over 11 days, comparable to the wild-type control. Day 1 to 11 post thaw; 10X magnification.

bit.bio-io1107-gfp-glutamatergic-neurons-gene-expression-rt-qpcr

GFP ioGlutamatergic Neurons demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following deterministic programming

Gene expression analysis demonstrates that GFP ioGlutamatergic Neurons (GFP) and wild-type ioGlutamatergic Neurons (WT) lack the expression of pluripotency marker OCT4 (POU5F1) at day 11, while robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic-specific (VGLUT1 and VGLUT2) markers, as well as the glutamate receptor GRIA4.

Gene expression levels were assessed by RT-qPCR, data normalised to HMBS; cDNA samples of the parental human iPSC line (iPSC) were included as reference. Data represents day 11 post-revival samples.

Co-culture workflow for GFP glutamatergic neurons and microglia

Easy-to-use co-culture protocol for GFP ioGlutamatergic Neurons with ioMicroglia

This protocol describes a method for co-culturing GFP ioGlutamatergic Neurons with ioMicroglia and associated disease models.

Vial limit exceeded

A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@bit.bio.

Human iPSC-derived glutamatergic neurons constitutively expressing GFP

GFP ioGlutamatergic Neurons are a fluorescent human excitatory neuronal model derived from our well-established wild-type ioGlutamatergic Neurons. These cells are engineered to constitutively express green fluorescent protein (GFP), and provide a powerful, ready-to-use tool for diverse applications.

Stable GFP expression enables easy, real-time tracking in complex, multi-cellular systems, facilitating the study of cellular interactions with glia, or network function when co-cultured with inhibitory neurons. The cells are ideal for live cell imaging for assessment of neurite outgrowth, neuronal morphology and survival in response to compound treatment.

GFP ioGlutamatergic Neurons are delivered cryopreserved and ready-to-culture. This eliminates the time and effort required to engineer your own GFP-expressing iPSC lines and manage complex directed differentiation protocols.

Benchtop benefits

Rapid results with ready-to-culture neurons stably expressing GFP

Live-cell imaging ready

Assess neurite outgrowth, neuronal morphology and survival in real-time.

Co-culture GFP glutamatergic neurons

Co-culture compatible

Easily track GFP glutamatergic neurons in complex culture with glia.

GFP glutamatergic neurons are easy-to-use

Easy-to-use

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

Cells arrive ready to plate


bit.bio_ioGlutamatergic_Neurons_WT_timeline

GFP 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 specifications

Download product brochure

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, age 55-60 years old (skin fibroblast),
Genotype APOE 3/4

Vial size

Small: >1 x 10⁶ viable cells,
Evaluation pack*: 3 small vials of >1 x 10⁶ viable cells

Quality control

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

Differentiation method

opti-ox deterministic cell programming

Recommended minimum seeding density

30,000 cells/cm²

User storage

LN2 or -150°C

Format

Cryopreserved cells

Product use

ioCells are for research use only

Applications

Live-cell imaging
Co-culture
High-content screening
Neurotoxicology
Drug discovery

* Evaluation packs are intended for first-time users, or for existing users testing a new cell type or derivative. A user can request multiple evaluation packs as long as each one is for a different product, with only one pack allowed per product.

ioGlutamatergic Neurons Customer Testimonials

An image of Dr Shushant Jain

Dr Shushant Jain

Group Leader | In Vitro Biology | Charles River, 2021

"These cells enable us to move rapidly as from the moment of plating within 4-7 days we have mature and functional neurons."

An image of Dr Mariangela Iovino

Dr Mariangela Iovino

Senior Group Leader | Biology Discovery | Charles River

“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.”

An image of Dr Koby Baranes

Dr Koby Baranes

Research Associate | University of Cambridge

"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.”

An image of Dr Jeremy Anton

Dr Jeremy Anton

Scientist | Charles River

"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."
Technical data

Highly characterised and defined

Flow cytometry analysis of GFP expression at day 11 and day 21

GFP expression by flow cytometry in GFP glutamatergic neurons

Flow cytometry analysis demonstrating GFP expression in >99% of cells for GFP ioGlutamatergic Neurons cultured until day 11 (centre), and no GFP expression seen in wild-type ioGlutamatergic Neurons (left).  At day 21, the percentage of cells expressing GFP has not decreased, indicating there is no silencing of the reporter gene (right). 

GFP ioGlutamatergic Neurons exhibit comparable expression of neuron-specific markers to the wild-type control

Protein expression for GFP glutamatergic neurons by ICC

Immunofluorescent staining 11 days post-revival shows similar homogenous expression of pan-neuronal markers MAP2 and TUBB3 (upper panel) and the glutamatergic transporter VGLUT2 (lower panel) in both GFP ioGlutamatergic Neurons and the wild-type control.

The GFP signal is visible exclusively in GFP ioGlutamatergic Neurons and absent in the wild-type control (lower panel). 10X magnification.

GFP ioGlutamatergic Neurons form structural neuronal networks by day 11

GFP glutamatergic neurons show expected neuronal morphology

GFP ioGlutamatergic Neurons mature rapidly, show glutamatergic neuron morphology and form structural neuronal networks over 11 days. Day 1 to 11 post thawing; 10X magnification.

GFP ioGlutamatergic Neurons demonstrate gene expression of neuronal-specific and glutamatergic-specific markers following deterministic programming

Gene expression in GFP glutamatergic neurons by RT-qPCR

Gene expression analysis demonstrates that GFP ioGlutamatergic Neurons (GFP) and wild-type ioGlutamatergic Neurons (WT) lack the expression of pluripotency marker OCT4 (POU5F1) at day 11, while robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic-specific (VGLUT1 and VGLUT2) markers, as well as the glutamate receptor GRIA4.

Gene expression levels were assessed by RT-qPCR, data normalised to HMBS; cDNA samples of the parental human iPSC line (iPSC) were included as reference. Data represents day 11 post-revival samples.

Co-culture protocol

Easy-to-use co-culture protocol for GFP ioGlutamatergic Neurons with ioMicroglia

Co-culture workflow for GFP glutamatergic neurons and microglia

This protocol describes a method for co-culturing GFP ioGlutamatergic Neurons with ioMicroglia and associated disease models.

View the co-culture protocol
Technical data

GFP glutamatergic neurons and microglia co-culture

Easy-to-use co-culture protocol for GFP ioGlutamatergic Neurons with ioMicroglia

Co-culture workflow for GFP glutamatergic neurons and microglia

This protocol describes a method for co-culturing GFP ioGlutamatergic Neurons with ioMicroglia and associated disease models.

View the co-culture protocol

How to culture ioGlutamatergic Neurons

In this video, our scientist will take you through the step-by-step process of how to thaw, seed and culture ioGlutamatergic Neurons.

Product resources

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CRISPRi-Ready ioGlutamatergic Neurons | User Manual User manual
CRISPRi-Ready ioGlutamatergic Neurons | User Manual
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CRISPRa-Ready ioGlutamatergic Neurons | User Manual User manual
CRISPRa-Ready ioGlutamatergic Neurons | User Manual
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bit.bio
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Human iPSC-Based Models of Glial Cells for Studying Neurodegenerative Disease Webinar
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ioGlutamatergic Neurons Wild Type and related disease models | User Manual User manual
ioGlutamatergic Neurons Wild Type and related disease models | User Manual

DOC-1289 4.0

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2025

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Running Large-Scale CRISPR Screens in Human Neurons Webinar
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Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming Webinar
Addressing the Reproducibility Crisis | Driving Genome-Wide Consistency in Cellular Reprogramming

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Rethinking Developmental Biology With Cellular Reprogramming Webinar
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Modelling neurodevelopment | Investigating the impact of maternal immune activation on neurodevelopment using human iPSC-derived cells Webinar
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Expand your research

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Co-culture human excitatory and inhibitory neurons to model neurodegenerative disease
Build functional excitatory - inhibitory in vitro cultures
Combine defined, consistent glutamatergic and GABAergic neurons
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CRISPR-ready hiPSC-derived cells ready for CRISPR screens
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Perform gene knockout, activation and interference in iPSC-derived neurons and glia
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CRISPR-ready hiPSC-derived cells ready for CRISPR screens
CRISPR-Ready ioCells stably express Cas9 nuclease or dCas9 variants and come with optimised cell culturing and guide delivery protocols.
Start measuring readouts from gene perturbations and CRISPR screens within days

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Multi-cell model for in vitro studies and disease modelling
Access our in vitro neuroscience toolkit to develop complex models for investigating cellular communication and disease mechanisms in the human context.

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