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ioGlutamatergic Neurons
PRKN R275W/R275W™

Human iPSC-derived Parkinson's disease model

ioGlutamatergic Neurons PRKN R275W/R275W are opti‑ox™ precision reprogrammed glutamatergic neurons carrying a genetically engineered homozygous 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 homozygous R275W mutation in Parkinson’s disease.

Related disease model cells are available now with a heterozygous PRKN R275W mutation, and both can be used alongside their genetically matched control, ioGlutamatergic Neurons™.

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Benchtop benefits

Make True Comparisons

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

Scalable

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

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.

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 cellular reprogramming

Recommended seeding density

30,000 cells/cm²

User storage

LN2 or -150°C

Format

Cryopreserved cells

Genetic modification

Homozygous 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

Highly characterised and defined

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

ICC showing that ioGlutamatergic Neurons PRKN R275W/R275W express pan neuronal markers

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/R275W compared to the isogenic control. 100X magnification.

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

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/R275W 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/R275W following reprogramming

ioGlutamatergic_neurons-PRKN-R275W-R275W-rt-qPCR-PRKN2

Gene expression analysis demonstrates that ioGlutamatergic Neurons PRKN R275W/R275W 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

PRKN R275W/R275W arrive ready to plate and are experiment ready in days

ioGlutamatergic Neurons PRKN R275W/R275W 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: Induction, which is carried out at bit.bio (Phase 0), Stabilisation for 4 days (Phase 1), and Maintenance (Phase 2) during which the ioGlutamatergic Neurons PRKN R275W/R275W mature. Phases 1 and 2 after revival of cells are carried out by the customer.

Industry leading seeding density

ioGlutamatergic Neurons PRKN R275W/R275W are compatible with plates ranging from 6 to 384 wells and are available in two vial sizes, tailored to suit your experimental needs with minimal waste.

The recommended seeding density is 30,000 cells/cm², compared to up to 500,000 cells/cm² for other available 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 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.

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Supporting documentation

User Manual

Statement of Use

Product resources

Talk

Precision Cellular Reprogramming for Scalable and Consistent Human Neurodegenerative Disease Models

Madeleine Garrett | Field Application Specialist | bit.bio

2023

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Poster

Development and characterisation of a robust in vitro disease model to study tauopathies

V1
Charles River & bit.bio

2022

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Poster

Validation of ALS-relevant phenotypes in precision reprogrammed iPSC-derived glutamatergic Neurons containing a TDP-43 M337V mutation.

V1
Charles River & bit.bio

2022

Download

Video

Partnering with Charles River to advance CNS drug discovery with ioGlutamatergic Neurons™

Charles River

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Video tutorial

Culturing Glutamatergic Neurons | How-to Video

Dr Kaiser Karim | Scientist
bit.bio

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Video tutorial

Preparing Culture Vessels for Glutamatergic Neurons | How-to Video

Dr Kaiser Karim | Scientist
bit.bio

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Video

In Conversation with Charles River

Dr Marijn Vlaming | Head of Biology
Charles River

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Addressing current challenges of in vitro cell models

Read this blog to find out how experts from across academia and industry are approaching the challenges of reproducibility of in vitro cell models as well as potential solutions.

Read the blog