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.
ioGlutamatergic Neurons PRKN R275W/R275W 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/R275W compared to the isogenic control. 100X magnification.
ioGlutamatergic Neurons PRKN R275W/R275W form structural neuronal networks by day 11
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
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
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
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 cells are compatible with plates ranging from 6 to 384 wells. The recommended seeding density is 30,000 cells/cm2, compared to up to 500,000 cells/cm2 for other similar products on the market. This means scientists are able to do more with every vial and expand experimental design within budget without losing out on quality. Resulting in more experimental conditions, more repeats, and more confidence in the data. 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.
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
Homozygous R275W missense mutation in the PRKN gene
Parkinson's disease research Drug discovery and development Disease modelling