Marker Expression: Spinal Motor Neurons (BX-0100) have high neuronal purity (>90%) and consist of greater than 70% motor neurons. Labeling with the pan-neuronal marker MAP2 (green) and the motor neuron-specific marker FOXP1 (red) highlight the purity of this neuronal product.
Morphology: Spinal Motor Neurons (BX-0100) are adherent and exhibit substantial neurite outgrowth within the first week in culture. Calcein staining (green) demonstrates the elaborate processes of the Spinal Motor Neurons in culture.
Function: Spinal Motor Neurons (BX-0100) exhibit pronounced electrophysiological activity after two weeks in culture, as demonstrated by multi-electrode array (MEA) recordings.
Recommended protocols developed by BrainXell utilize proprietary supplements that are included with each purchase of neurons. The recommended protocol will depend on the application. Contact BrainXell application scientists to learn more.
Calcium Influx Assays:
Changes in calcium concentration are closely tied to neuronal activity as action potentials are associated with large pre-synaptic calcium influx and a notable rise in postsynaptic calcium at excitatory synapses. This can be observed experimentally by stimulating the neurons or culturing the neurons under suitable conditions to form mature networks that exhibit spontaneous oscillations. The influx of calcium can be measured using a variety of calcium-sensitive fluorescent dyes, which are commercially available.
Spinal Motor Neurons (BX-0100) were cultured in 96-well plates for three weeks and then loaded with Calbryte-520 (AAT Bioquest). Spontaneous oscillations were recorded in all wells simultaneously using an FDSS/µCell Functional Drug Screening System (Hamamatsu).
Multi-electrode arrays (MEA) measure extracellular voltage changes that occur as neurons fire action potentials. These measurements reveal the firing patterns of individual neurons as well as the patterns of neuronal networks that exist in the cell culture. Such measurements are non-invasive and allow for repeated recordings.
Spinal Motor Neurons (BX-0100) were cultured on Axion Biosystems MEA plates for several weeks and recorded regularly starting at one week. Below, a time course of the number of active electrodes, mean firing frequency, and synchrony index reveal the development of neuronal activity in the spinal motor neurons over several weeks in culture.
The raster plot of spike activity shows network bursting observed on day 20 in culture. The resulting activity can be analyzed for various parameters of the spiking patterns.
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