Marker Expression: Mixed Cortical Neurons (BX-0500) have high neuronal purity (>90%) and consist of glutamatergic and GABAergic neurons in an 80:20 ratio. Labeling with the pan-neuronal marker MAP2 (green), the glutamatergic-specific marker CTIP2 (purple), and GABA (red) highlight the purity and composition of this neuronal product.
Morphology: Mixed Cortical Neurons (BX-0500) are adherent cells and exhibit substantial neurite outgrowth within a week in culture. Calcein staining (green) demonstrates the characteristic cell shape and extension of processes of cortical neurons in culture.
Function: Mixed Cortical Neurons (BX-0500) 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.
Mixed Cortical Neurons (BX-0500) 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.
Mixed Cortical Neurons (BX-0500) were co-cultured with astrocytes on Axion Biosystems MEA plates for several weeks and recorded regularly from two to four weeks. Below, a time course of the number of active electrodes, mean firing frequency, and synchrony index reveal the development of neuronal activity in the mixed cortical neurons over several weeks in culture.
The example raster plot of spike activity shows network bursting observed on day 21 for Mixed Cortical Neurons cultured alone. The resulting activity can be analyzed for various parameters of the spiking patterns.
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