ioNEURONS/glut

Human-Induced Glutamatergic Neurons

ioNEURONS/glut cells have been reprogrammed from human induced pluripotent stem cells (iPSC) using our precise reprogramming technology:
opti-ox¹
(optimized inducible overexpression). Human stem cells, within days, convert into consistent, mature, functional glutamatergic neurons providing a high quality human model for the study of neurological activity and disease.

ioNEURONS/glut cultures consist mainly of glutamatergic neurons (>80%) characterised by the expression of the glutamate transporter genes VGLUT1 and VGLUT2. The minor remaining fraction of the neuronal population express marker genes of cholinergic neurons. A bulk RNA-seq analysis shows that ioNEURONS/glut have a rostral CNS identity and express the classical cortical marker genes FOXG1 and TBR1 (data not shown).

Ready-to-culture cells are suitable as models for research in cell-type specific biology, target validation and drug screening in pharmaceutical R&D, and toxicology testing.

¹ Pawlowski et al., Stem Cell Report 2017

Advantages:

– Highly defined and well characterized
– Fast maturation
– Early electrical signal
– Easy culturing
– Production specification


Product applications:

Academic research
– Drug development
– Neurotoxicology
– High-throughput screening
– Genetic screening (e.g. CRISPR screening)

For more information please email:
info@bit.bio

Batch to batch reproducibility and homogeneity create a stable human model for excitatory neuronal activity and disease.

Ready for experimentation as early as 2 days post revival and form functional neuronal networks at 17 days.

Industrial scale quantities at a price point that allows the cells to be used from research to screening scale.

Cells arrive programmed to rapidly mature upon revival. One medium required in a two-step protocol.

ioNEURONS/glut express glutamatergic neuron-specific markers

TUBB3

VGLUT1

MAP2

VGLUT2

Immunofluorescent staining on post-revival day 11 demonstrates homogenous expression of pan-neuronal proteins (MAP2 and TUBB3) and glutamatergic neuron-specific transporters (VGLUT1 and VGLUT2). Cells exhibit neurite outgrowth.

ioNEURONS/glut after revival over the course of the first 11 days

Day 1 to 11 post-thawing; 400X magnification; scale bar: 100µm.

ioNEURONS/glut display neuronal activity that matures over-time

Examples of MaxOne high-resolution multi electrode array (MEA) recordings of ioNEURONS/glut in BrainPhys™ media. The activity maps show firing rate (A), spike amplitude (B) and % of active electrodes (C). Results demonstrate a time-dependent increase of spontaneous activity during neuronal maturation from 2 to 3 weeks post-revival.

Iovino et al., Molecular and functional characterization of stem cells-derived glutamatergic neurons ioNEURONS/glut in support of drug discovery applications., Charles River Laboratories.

ioNEURONS/glut show good suitability for high-throughput screening in 384-well format plates

Cytotoxicity CellTiter-Glo®️ (CTG) and TR-FRET (HTRF®️) assays for AKT serine/threonine kinase 1 (AKT) and Huntingtin (HTT) proteins were performed on ioNEURONS/glut in 384-well plates treated with tool compound (cmp) at day 9 post-revival. Compound titration results in a concentration response curve for all three assays (mean±sd of 2 replicates). CTG assay on ioNEURONS/glut shows an excellent average signal/ background ratio and high suitability for HTS. HTRF® assays on ioNEURONS/glut show lower signals but with low variability, and could therefore also provide a suitable platform for HTS.

Iovino et al., Molecular and functional characterization of stem cells-derived glutamatergic neurons ioNEURONS/glut in support of drug discovery applications., Charles River Laboratories. 

Arrive ready to plate

ioNEURONS/glut 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: 1. Induction (carried out at bit.bio) 2. Stabilization for 4 days with Dox 3. Maintenance during which the neurons mature.

Cost effective and flexible

ioNEURONS/glut SKUs

ioNEURONS/glut 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. Recommended seeding density for ioNEURONS/glut is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other available products on the market. One Small vial can plate a minimum of 0.5 x 24-well plate, 0.75 x 96-well plate, or 1 x 384-well plate. One Large vial can plate a minimum of 1 x 24-well plate, 1.5 x 96-well plate, or 2 x 384-well plates.

Product Information


Catalogue number: e001 Manufacture site: United Kingdom Sizes: Small & Large Product use: These cells are for research use only. It is not approved for human or animal use, or for application in in vitro diagnostic procedures Storage: Upon receiving, directly and immediately transfer the cells from dry ice to liquid nitrogen and keep the cells in liquid nitrogen until they are needed for experiments. Shipping info: Dry Ice Note: Limited Use License | Statement of Use