Sensory Neuron Maturation Maximizer Supplement

Sensory Neuron Maturation Maximizer Supplement

MATURATION MAXIMIZER is a fully defined medium supplement for enhancing the existing Sensory Neuron Maintenance Medium (ax0060). It works by mimicking in vivo signals between sensory neurons and their supporting cells. The supplement contains signaling factors present in the peripheral nervous system and in particular the native environment of sensory neurons. Utilizing this supplement accelerates the maturation of iPSC-derived sensory neurons in vitro.

iPSC-derived sensory neurons treated with the Maximizer express key markers, such as TRPv1, TRPa1, TRPv2, TRPv3 and TRPm8. Kinetic states of e.g. TRPv1 are in a fast response inward rectifying state, to agonists such as Capsaicin. Neuronal responses to capsaicin occur at 1μM for 90% of the sensory neuron population.

The MATURATION MAXIMIZER supplement makes the Human iPSC-derived sensory neuronal culture a more physiological relevant model.

Figure 1. Calcium Imaging of the iPSC-sensory neurons’ responses to 1 μM Capsaicin when cultured with and without Maximizer. When cultured with Sensory Neuron Maturation Maximizer Supplement, sensory neurons respond to low concentrations of Capsaicin indicating the presence of functional TRPV1 channels.

A. Average deltaF/Fo values for the unsupplemented and Maximizer-supplemented sensory neurons treated with vehicle control and 1 μM capsaicin on Day 20 of culture.

B & C. Pie Chart and Table showing capsaicin threshold concentrations for sensory neurons treated with and without Maximizer. 91% of Regions-Of-Interest (ROIs) show a response to the lowest concentration of capsaicin tested (1 μM) compared to only 19% of control cells and while 99% of the ROIs of treated sensory neurons respond to the highest concentration of capsaicin (100 μM), 57% of non-treated sensory neurons remain non-responsive.

This shows that even by Day 20, the Maximizer-treated sensory neurons show enhanced capsaicin responses compared to non-treated.

Fig 3. Electrophysiology of Axol’s Sensory Neurons when cultured with Sensory Neuron Maturation Maximiser Supplement

Representative MEA traces of sensory neurons with and without treatment of Maximizer. All traces are 60s and compare the same electrode in each figure.

A. Spontaneous activity. Treated cells showed significant spontaneous activity by Day 21 (top) compared to Day 28 (bottom)

B. Response to capsaicin on Day 22. Rising concentrations of capsaicin produced significantly increased activity in treated cells with minimal effect on untreated. This indicates the presence of functional TRPV1 channels in the treated sensory neurons

C. Response to menthol on Day 27. Rising concentrations of capsaicin produced significantly increased activity in treated cells with minimal effect on untreated. This indicates the presence of functional TRPM8 channels in the treated sensory neurons