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Human iPSC-Derived Motor Neuron

Motor Neuron Progenitors

Axol’s human iPSC-derived motor neuron progenitors have been differentiated using a combination of small molecules to regulate multiple signalling pathways to generate a near pure population of motor neurons progenitors which matured into an enriched population of mature motor neurons.

To support the recovery and maintenance of Axol’s human iPSC-derived motor neuron progenitors, we offer our motor neuron recovery medium which supports cell recovery after thawing and initial cell culture and our motor neuron maintenance medium which promotes the optimal growth and maturation of progenitors to iPSC-derived motor neurons.

We have developed human iPSC-derived motor neuron progenitors from both healthy and patient donors, which can be cultured in vitro to provide a physiologically relevant alternative to conventional animal-derived models for use in proof-of principle drug-screening assays and for modelling motor neuron degeneration.


Human iPSC-derived motor neuron progenitors

Cat. No. Product Name Starting Material Quantity
ax0078 Motor Neuron Progenitors (Healthy) Fibroblasts (74 yr Male) ≥2 x 106 cells
ax0073 Motor Neuron Progenitors (Healthy - sibling (C9ORF72 extension)) Fibroblasts (62 yr Male) ≥2 x 106 cells
ax0074 Motor Neuron Progenitors (ALS (C9ORF72 extension)) – coming soon! Fibroblasts (64 yr Female) ≥2 x 106 cells
ax0178 Human iPSC-Derived Motor Neuron Progenitor Kit (Male) Fibroblasts (74 yr Male) 1 Kit (≥2 x 106 cells, medium, coating reagents & growth factors)

Motor neuron culture media and reagents

Cat. No. Product Name Product Description Quantity
ax0071 Motor Neuron Recovery Medium Promotes viability of motor neuron progenitors 50 mL
ax0072 Motor Neuron Maintenance Medium For the maintenance of motor neurons 200 mL
ax0041 SureBond Coating solution providing an optimal surface for feeder-free growth & adherence 3 vials of SureBond (120 μL/vial)
ax0052 SureBond+ReadySet Coating reagents required for endpoint assays on glass 3 vials of SureBond (120 μL/vial) and 2 vials of ReadySet (10 mL/vial)
ax0044 Unlock Fully defined & gentle detachment buffer 25 mL

Phase contrast

Phase contrast images show the morphology of human iPSC-derived motor neuron progenitors upon recovery from thawing.

Phase contrast images of Axol Human iPSC-derived motor neuron progenitors at days 1, 3 and 5 after thawing. Human iPSC-derived motor neuron progenitors are seeded using motor neuron recovery medium into T-25 flasks (coated with SureBond) at a density of 100,000 cells/cm2. During initial recovery phase the motor neuron progenitors will grow as aggregated clumps as they proliferate and expand at least 3 fold. Images provided by Lucia Marani and Prof. Mark Lewis (Loughborough University).


Immunocytochemistry

Axol’s Human iPSC-Derived Motor Neuron Progenitors express cell fate markers.

Protein expression of OLIG2, nestin and SOX1 was confirmed in Axol iPSC-Derived Motor Neuron Progenitors using immunocytochemistry. iPSC-Motor Neuron Progenitors were cultured in motor neuron recovery medium (ax0071) for 3 days after thawing before staining for OLIG2, nestin and SOX1. nestin and SOX1 staining (red) confirmed early neural ectoderm marker and OLIG2 staining (red) confirmed motor neuron progenitor differentiation. Images were counterstained with Hoechst.

Axol’s human iPSC-derived motor neurons show the expression of HB9, LIM3 and ChAT, which are typically expressed in mature motor neurons.

Protein expression of HB9, LIM3 and ChAT was confirmed in Axol iPSC-derived motor neuron using immunocytochemistry. Human iPSC-motor neurons were cultured in motor neuron maintenance medium (ax0072) for 14 days post passaging before staining for motor neuron markers LIM3 (green), HB9 (red) and ChAT (red) along with staining for neural markers TUJ1 (green) and MAP2 (red). Merged images were counterstained with Hoechst.

Human iPSC-derived Motor Neurons cultured on glass coverslips coated with SureBond+ReadySet (ax0052) for 3 weeks show expression of the mature motor neuron marker SMI-32 (yellow) (neurofilament H non phosphorylated), nuclei were stained with DAPI (blue). Data kindly provided by Amit K. Chouhan, Abby Scurfield, and Dr Gareth Miles (University of St Andrews).


Electrical activity - whole cell patch clamp

Whole cell patch clamp recordings from human iPSC-derived motor neurons reveal the electrical properties and functional connectivity of our motor neurons.

Human iPSC-derived motor neurons cultured for A) 3 and B) 7 weeks show typical morphology when imaged using Infrared – differential interference contrast (IR-DIC) microscopy. B) IR-DIC image shows electrode used for voltage and current clamp recordings. Data kindly provided by Amit K. Chouhan and Dr Gareth Miles (University of St Andrews).

Voltage Clamp

Axol’s Human iPSC-derived Motor Neurons show synaptic activity at 7 weeks in culture.

(Ai) Synaptic activity recorded from a Human iPSC-derived Motor Neuron using whole-cell patch-clamp technique (voltage clamp mode). (Aii) Synaptic activity shown on a faster time scale to reveal the kinetics of individual postsynaptic currents. (B) Traces showing fast, inactivating Na+ currents recorded in response to brief depolarizing voltage steps. (C) Traces showing persistent K+ currents recorded in response to longer duration depolarizing voltage steps. Data kindly provided by Amit K. Chouhan and Dr Gareth Miles (University of St Andrews).

Current Clamp

Axol’s Human iPSC-derived Motor Neurons exhibits both single and repetitive firing of action potentials upon stimulation.

Human iPSC-derived Motor Neurons were cultured for 7 weeks on SureBond+ReadySet (ax0052) coated glass coverslips (A) Trace shows a single action potential recorded from a functional iPSC-derived Motor Neuron in response to brief current injection in current clamp mode. (G) Trace shows repetitive firing of action potentials recorded from a functional iPSC-derived Motor Neuron upon step current pulse injection. (H) A train of action potentials (10Hz) elicited in an iPSC-derived Motor Neuron by a series of depolarizing current pulses. Data kindly provided by Amit K. Chouhan and Dr Gareth Miles (University of St Andrews).


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