ioMotor Neurons have been precision reprogrammed from human induced pluripotent stem cells (iPSC) using opti-ox™ technology. Within days, cells convert consistently to defined, functional motor neurons, showing the expression of key lower motor neuron marker genes MNX1(HB9), FOXP1, ISL2 and cholinergic markers CHAT & SLC18A3 (VAChT) by day 4.
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ioMotor Neurons have been extensively characterised, indicating a spinal motor neuron identity (cervical region) confirmed through FOXP1, ISL2, MNX1 and HOX gene expression that is measurable just 4 days post-revival. From day 14, >80% cells express MNX1 and show functional neuronal activity in astrocyte co-culture that is in line with primary motor neurons cultured in similar conditions. Functional activity was also evaluated in monoculture, demonstrating no significant activity, as expected; indicating that the cells form a highly pure and homogenous neuronal population unable to undertake synaptogenesis without glial cells.
ioMotor Neurons represent an accurate in vitro model of lower motor neurons (indicated spinal – cervical region identity), enabling scientists to build physiological relevance into their experiments at scales from single cell analysis to high content imaging, helping bridge translational gaps in motor neuron disease research and neurotoxicology.
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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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ioMotor Neurons acquire a rapid motor neuronal phenotype, without clumping
Time-lapse video capturing the rapid and homogeneous motor neuronal phenotype acquisition upon thawing of cryopreserved ioMotor Neurons, showing no signs of clumping. 11 day time course.
ioMotor Neurons form a homogenous neuronal network by day 4
ioMotor Neurons mature rapidly and form homogenous populations over 18 days. Day 1 to 18 post thawing; 100X magnification.
Rapid gain of functional activity
Functional neuronal networks are detected in astrocyte co-culture from day 14
ioMotor Neurons are functional – showing activity in astrocyte co-culture that increases over time as networks mature. Mean firing rates (electrode spike count divided by the total time of the recording period) is shown to increase substantially throughout the course of the experiment, as demonstrated by multielectrode array activity (MEA).
Spontaneous neuronal activity is exhibited from as early as day 14 and continues to increase up to the final measured timepoint, day 42.
Highly characterised and defined
Immunocytochemistry shows protein expression of key motor neuron markers
Immunofluorescent staining on post-revival day 4 and day 11 demonstrates homogenous expression of the pan-neuronal protein TUBB3, motor neuron specific marker ISL2, the cholinergic marker ChAT and nuclear staining (DAPI).
Immunofluorescent staining on post-revival day 4 and day 11 demonstrates homogenous expression of the pan-neuronal protein MAP2, motor neuron specific marker HB9, the cholinergic marker VAChT and nuclear staining (DAPI).
RT-qPCR shows gene expression of key motor neuron markers
RT-qPCR gene expression on post-revival days 1, 4, 11 & 18 demonstrates rapid acquisition of motor neuron genotype, shown by the expression of pan-neuronal, cholinergic & key lower motor neuron markers from as early as day 1. Pluripotency markers NANOG and OCT4 are swiftly downregulated.
Bulk RNA-sequencing exhibits a HOX gene signature indicative of a spinal motor neuron (cervical region) identity
Expression of HOX genes was evaluated using bulk RNA sequencing data. This heatmap shows expression of genes from the B cluster and expression of HOXC4 and HOXC5, although at lower levels. This data, together with the marker expression from single cell RNA sequencing, suggests that ioMotor Neurons have a spinal cord (cervical region) identity. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single cell RNA-sequencing shows ioMotor Neurons form a pure population (>99.9%) of neurons
Single cell RNA-sequencing analysis was performed with ioMotor Neurons at four timepoints: day 0 (iPSCs), 4, 7, and 14. Gene expression was assessed by 10x Genomics single cell RNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells. By day 14, the population has a distinct expression profile indicating a pure population (>99.9%) of post-mitotic neurons, demonstrated through the expression pan-neuronal markers MAP2 and TUBB3.
Single cell RNA-sequencing shows ioMotor Neurons express key spinal motor neuron markers, >80% of cells express MNX1 on day 14
Starting from day 4, the expression of the key spinal motor neuron marker genes MNX1 (HB9), FOXP1, and ISL2 is detected in the culture, with >80% of cells expressing MNX1 on day 14. These percentages are likely to be an underestimation due to limitation of single cell RNA sequencing, as ICC for HB9 & ISL2 shows homogeneous expression of these markers in our cultures
Single cell RNA-sequencing shows a high proportion of ioMotor Neurons express cholinergic markers by day 7
Within 7 days, the expression of the key cholinergic marker genes CHAT & SLC18A3 (VAChT) are detected in a high proportion of ioMotor Neurons.
Bulk RNA-sequencing demonstrates high batch-to-batch consistency of ioMotor Neurons
Bulk RNA sequencing analysis was performed on three independent batches of ioMotor Neurons at three different time points throughout the reprogramming protocol. Principal component analysis represents the variance in gene expression between the batches of ioMotor Neurons. This analysis shows high consistency between each batch of ioMotor Neurons at each given timepoint. Populations of ioMotor Neurons with equivalent expression profiles can be generated consistently from every vial, allowing confidence in experimental reproducibility. Note, this data is from cells in continuous culture and not cryopreserved cells.
Industry leading seeding density
Do more with every vial
The seeding density of our human iPSC-derived motor neurons has been optimised and validated to a recommended seeding density of 30,000 cells/cm2. This means scientists can 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.
Cell arrive ready to plate
ioMotor Neurons 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. Stabilisation for 2 days. 2. Pre-maintenance for an additional 2 days. 3. Maintenance of cells according to the protocol and recommended media for the duration of assay requirements.
Human iPSC line
6, 12, 24, 96 and 384 well plates
Caucasian adult male (skin fibroblast)
Small: >1 x 10⁶ viable cells
Sterility, protein expression (ICC) and gene expression (RT-qPCR)
opti-ox cellular reprogramming
Recommended seeding density
LN2 or -150°C
ioCells are for research use only
Neurodegeneration research ALS disease modelling Electrophysiological analysis Drug development & discovery Neuromuscular research Neurotoxicology