cat no | io1027

ioMotor Neurons

Human iPSC-derived motor neurons

ioMotor Neurons from human induced pluripotent stem cells (iPSC) deterministically programmed 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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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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Benchtop benefits

Functional

Functional neuronal networks are detected in co-culture with astrocytes from day 14.

Quick and easy

Within 4 days post revival cells are ready for experimentation, displaying motor neuronal morphology without clumping.

Defined

>80% cells express key lower motor neuron markers indicating a spinal motor neuron identity (cervical region). >99.9% neuronal population.

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

iomotor_neuron_gene_expression_RTqPCR_data

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.

Batch-to-batch consistency

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 ioMotor Neurons and related disease models 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.

Ready within days

Schematic overview of the timeline in the user manual

ioMotor Neurons and related disease models are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media.

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