cat no | io1107 Early Access
GFP ioGlutamatergic Neurons
Human iPSC-derived glutamatergic neurons constitutively expressing GFP
- Cryopreserved human iPSC-derived cells powered by opti-ox that are ready for experiments in days
- Glutamatergic neurons constitutively expressing GFP, easy to track in multi-cell cultures, ideal for live-cell imaging
- Co-culture compatible with ioMicroglia and ioAstrocytes
Human iPSC-derived glutamatergic neurons constitutively expressing GFP
ioGlutamatergic Neurons Media Kit
Cell culture media kit for the culture of ioGlutamatergic Neurons up to 14 days post-thaw
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.
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 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.
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.
View the step-by-step RNA extraction and RT-qPCR protocol used to generate this data
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.
Phenotypic characterisation of a human iPSC-derived tri-culture using ioGlutamatergic Neurons, ioAstrocytes, and ioMicroglia
Using our fully optimised protocol, ioGlutamatergic Neurons (MAP2, red), ioMicroglia (IBA1, yellow) and ioAstrocytes (vimentin, cyan) were co-cultured to create a highly defined CNS model. High-resolution ICC analysis confirms the successful co-localisation and morphological health of three distinct cell types within a unified environment. By day 7, the protocol yields a highly consistent, integrated network suitable for complex cell modelling. DAPI (blue) highlights the total cell density and integrity of the culture. This protocol is compatible with derivative products of the three cell types, ensuring straightforward implementation across experimental workflows.
Efficient mRNA transfection into ioGlutamatergic Neurons
ioGlutamatergic Neurons are efficiently transfected and show sustained long-term expression of mRNA encoding GFP. ioGlutamatergic Neurons were imaged from day 1 post-thaw and throughout the experiment to assess transfection efficiency and evaluate potential cytotoxic effects of the transfection protocol. Day 1 images were captured prior to transfection on the same day.
Download the step-by-step protocol for lipid-based delivery of synthetic mRNA into ioGlutamatergic Neurons.
Lipid-based delivery of synthetic mRNA into ioGlutamatergic Neurons
ioGlutamatergic Neurons were transfected 24 hours post-thaw using Lipofectamine™ Stem Transfection Reagent. The transfection efficiency was evaluated by fluorescence imaging over 18 days after mRNA delivery, resulting in high transfection efficiency (close to 100%) and long-term sustained GFP expression.
Quantification of the GFP signal shows a decrease in GFP intensity over time, while the percentage of GFP-positive cells remains largely unchanged over time.
(A) The percentage of GFP-positive cells from two independent experiments.
(B) GFP intensity, quantified in successfully transfected cells from two independent experiments is quantified and normalised to day 2 (24 hours post-transfection).
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.
Benchtop benefits
Live-cell imaging ready
Assess neurite outgrowth, neuronal morphology and survival in real-time.
Co-culture compatible
Easily track GFP glutamatergic neurons in complex culture with glia.
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
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
ioGlutamatergic Neurons Customer Testimonials
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."
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.”
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.”
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."
Highly characterised and defined
Flow cytometry analysis of GFP expression 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 (right).
GFP ioGlutamatergic Neurons exhibit comparable expression of neuron-specific markers to the wild-type control
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 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 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.
View the step-by-step RNA extraction and RT-qPCR protocol used to generate this data
Co-culture protocol
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.
Tri-culture protocol
Phenotypic characterisation of a human iPSC-derived tri-culture using ioGlutamatergic Neurons, ioAstrocytes, and ioMicroglia
Using our fully optimised protocol, ioGlutamatergic Neurons (MAP2, red), ioMicroglia (IBA1, yellow) and ioAstrocytes (vimentin, cyan) were co-cultured to create a highly defined CNS model. High-resolution ICC analysis confirms the successful co-localisation and morphological health of three distinct cell types within a unified environment. By day 7, the protocol yields a highly consistent, integrated network suitable for complex cell modelling. DAPI (blue) highlights the total cell density and integrity of the culture. This protocol is compatible with derivative products of the three cell types, ensuring straightforward implementation across experimental workflows.
GFP glutamatergic neurons and microglia co-culture
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.
Tri-culture with ioAstrocytes and ioMicroglia
Phenotypic characterisation of a human iPSC-derived tri-culture using ioGlutamatergic Neurons, ioAstrocytes, and ioMicroglia
Using our fully optimised protocol, ioGlutamatergic Neurons (MAP2, red), ioMicroglia (IBA1, yellow) and ioAstrocytes (vimentin, cyan) were co-cultured to create a highly defined CNS model. High-resolution ICC analysis confirms the successful co-localisation and morphological health of three distinct cell types within a unified environment. By day 7, the protocol yields a highly consistent, integrated network suitable for complex cell modelling. DAPI (blue) highlights the total cell density and integrity of the culture. This protocol is compatible with derivative products of the three cell types, ensuring straightforward implementation across experimental workflows.
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