axoCells™ motor neuron maintenance media, 200 ml

Phenotypic characterization: morphology

axoCells motor neurons demonstrate expected physiological morphology with uniform cell clusters and large cabling neuronal networks.

Phase contrast images of axoCells motor neurons matured from progenitors over 21 days.

Phenotypic characterization: ICC

axoCells motor neurons express key markers (TUJ1, ChAT) demonstrating phenotypic relevance.

Immunocytochemistry of day 21 mature axoCells motor neurons demonstrating presence of key markers (ChAT and TUJ1). Images captured on a Leica microscope x20 magnification. Error bar= 200um

Functional relevance: MEA

axoCells motor neurons exhibit synchronized burst firing, demonstrating their functional relevance

axoCells motor neuron firing assessed at day 10 using the Axion Maestro Pro multi-electrode array (MEA) system. Here you can see sodium spike firing and network burst firing responses of the motor neurons.

Left: sodium spike profile, right: Raster plot showing burst firing events (blue boxes) with synchronized firing highlighted in pink boxes.

Functional relevance: SNA (spontaneous neural activity)

axoCells motor neurons exhibit regular, synchronized firing on calcium imaging, demonstrating their functional relevance.

Firing pattern of axoCells motor neurons at day 21, transfected with NeuroBurst (calcium-sensitive lentivirus driven off the synapsin reporter) to pick up spontaneous neuronal activity on an IncuCyte. This demonstrates regular, synchronized firing with a mean correlation of 0.92. Potential parameters measured include mean correlation, burst rate, burst duration and burst strength.

Disease line vs control: ALS phase contrast

ALS-derived motor neurons form smaller, less uniform bodies with thinner neurite extensions compared to healthy axoCells motor neurons.

Phase contrast images of mature axoCells motor neurons (day 21) from healthy (left, ax0078) and ALS (right, ax0074) background.

Disease line vs control: ALS ICC

Compared to healthy axoCells motor neurons, ALS neurons form smaller, less uniform bundles, with thinner neurites. Healthy axoCells motor neurons form large, thick cabling between uniformly clustered bundles.

Immunocytochemistry of healthy control-derived axoCells motor neurons (top) and ALS-derived axoCells motor neurons (bottom) at day 21. Composite image for each channel and merged. Image captured on a Leica microscope at x20 magnification

Disease line vs control: SNA

Compared to healthy axoCells motor neurons, ALS-derived motor neurons exhibit less frequent and less-synchronized firing.

axoCells healthy control and ALS-derived motor neurons were matured for 21 days and transfected with Neuroburst Orange to look at spontaneous neuronal activity on the IncuCyte S3. Healthy motor neurons show regular synchronous firing compared to ALS which fire more frequently, for shorter duration and in a less-synchronized manner (shown by burst duration and rate). N=3, ***p<0.001, ****p<0.0001

axoModels: neuromuscular junction (NMJ)

Here is an example of a complex model we have built with motor neurons. As part of our axoModels offering, we are collaborating with biopharma organizations to build complex in vitro models, including an NMJ.

axoCells motor neurons matured on a microfluidics device with skeletal muscle in the opposite chamber. Neurite extensions can be observed passing through the grooves and innervating the skeletal muscle to form functional neuromuscular junctions (shown by bungarotoxin staining). A 3D model (not shown) has also been developed using a scaffold for the motor neurons, which are placed on top of skeletal muscle.