io1013S-ioGlutamatergic-Neurons-PRKN-R275W-heterozygous-ICC-MAP2 compressed

cat no | io1013

ioGlutamatergic Neurons
PRKN R275W/WT

Human iPSC-derived Parkinson's disease model

ioGlutamatergic Neurons PRKN R275W/WT are opti‑ox deterministically programmed glutamatergic neurons carrying a genetically engineered heterozygous mutation in the PRKN gene encoding the Parkin protein. These cells offer a rapidly maturing, disease relevant and isogenic system for investigating the molecular and cellular significance of a heterozygous R275W mutation in Parkinson’s disease.

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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.

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Benchtop benefits

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Pair the ioDisease Model Cells with the genetically matched wild-type ioGlutamatergic Neurons to directly investigate the effect of heterozygous expression of mutant Parkin protein on disease.

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Scalable

With opti-ox technology, we can make billions of consistently programmed cells, surpassing the demands of industrial workflows.

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Quick

The disease model cells and isogenic control are experiment ready as early as 2 days post revival, and form structural neuronal networks at 11 days.

Technical data
Technical data
Product information Supporting documentation Resources Related products

Highly characterised and defined

ioGlutamatergic Neurons PRKN R275W/WT express neuron-specific markers comparably to the isogenic control

ICC showing glutamatergic neurons with heterozygous R275W mutation in the PRKN gene expressing pan-neuronal markers in an equivalent manner to an isogenic control

Immunofluorescent staining on post-revival day 11 demonstrates similar homogenous expression of pan-neuronal proteins TUBB3 and MAP2 (upper panel) and glutamatergic neuron-specific transporter VGLUT2 (lower panel) in ioGlutamatergic Neurons PRKN R275W/WT compared to the isogenic control. 100X magnification.

ioGlutamatergic Neurons PRKN R275W/WT form structural neuronal networks by day 11

Incucyte imaging of ioGlutamatergic Neurons PRKN R275W/WT over 11 days, showing the cells form structural neuronal networks

ioGlutamatergic Neurons PRKN R275W/WT mature rapidly and form structural neuronal networks over 11 days, when compared to the isogenic control. Day 1 to 11 post thawing; 100X magnification.

ioGlutamatergic Neurons PRKN R275W/WT demonstrate gene expression of neuronal and glutamatergic-specific markers following deterministic programming

RT-qPCR of ioGlutamatergic Neurons PRKN R275W hom showing expression of pan neuronal and glutamatergic markers

Gene expression analysis demonstrates that ioGlutamatergic Neurons PRKN R275W/WT and the isogenic control (WT) lack the expression of pluripotency markers (NANOG and OCT4) at day 11, whilst 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 expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.

Disease-related PRKN is expressed in ioGlutamatergic Neurons PRKN R275W/WT following deterministic programming

RT-qPCR of ioGlutamatergic Neurons PRKN R275W/WT showing expression of the PRKN gene

Gene expression analysis demonstrates that ioGlutamatergic Neurons PRKN R275W/WT and the isogenic control (WT) express the PRKN gene encoding the Parkin protein. Gene expression levels were assessed by RT-qPCR (data expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.

Cells arrive ready to plate

ioGlutamatergic_Neurons_and_disease_models_timeline

ioGlutamatergic Neurons PRKN R275W/WT 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: Phase 1, Stabilisation for 4 days; Phase 2, Maintenance, during which the neurons mature. Phases 1 and 2 after revival of cells are carried out by the customer.

Industry leading seeding density

ioGlutamatergic_Neurons_seeding_density_small_96_384

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. This means every vial goes further, enabling more experimental conditions and more repeats, resulting in more confidence in the data.

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 106 viable cells

Quality control

Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)

Differentiation method

opti-ox deterministic cell programming

Recommended seeding density

30,000 cells/cm2

User storage

LN2 or -150°C

Format

Cryopreserved cells

Genetic modification

Heterozygous R275W missense mutation in the PRKN gene

Applications

Parkinson's disease research
Drug discovery and development
Disease modelling

Product use

ioCells are for research use only

Product resources

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Ferreira | Senior Product Manager | bit.bio, date_published=1732233600000, sort_date=1732233600000, tags=[{label=ioGlutamatergic Neurons, value=ioGlutamatergic Neurons}, {label=ioOligodendrocyte-like cells, value=ioOligodendrocyte-like cells}], media_contact=null, listing_button_label=Watch now}])
Human iPSC-Based Models of Glial Cells for Studying Neurodegenerative Disease Webinar
Human iPSC-Based Models of Glial Cells for Studying Neurodegenerative Disease
Valentina Fossati, PhD | Senior Research Investigator | The New York Stem Cell Foundation

Inês Ferreira | Senior Product Manager | bit.bio
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ioGlutamatergic Neurons Brochure
ioGlutamatergic Neurons

bit.bio

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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

V11

bit.bio

2024

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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
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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 

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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

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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

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

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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.

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