ioGlutamatergic Neurons PRKN R275W/WT are opti‑ox™ precision reprogrammed 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.
ioGlutamatergic Neurons PRKN R275W/WT express neuron-specific markers comparably to the 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
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 reprogramming
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 reprogramming
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 PRKN R275W/WT 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/WT mature. Phases 1 and 2 after revival of cells are carried out by the customer.
Industry leading seeding density
ioGlutamatergic Neurons PRKN R275W/WT 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.
Human iPSC line
Normal (46, XY)
6, 12, 24, 48, 96 & 384 well plates
Caucasian adult male (skin fibroblast)
Small: >1 x 106 viable cells
Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)
opti-ox cellular reprogramming
Recommended seeding density
LN2 or -150°C
Heterozygous R275W missense mutation in the PRKN gene
Parkinson's disease research Drug discovery and development Disease modelling