Neural Circuit and Innervation

Neural co-culture in 6 well Cytoview MEA plate with ibidi insert
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The brain is a collection of individual, but interconnected functional circuits.  Additionally, circuits within the nervous system interact with various other biological systems in the human body. Compartmentalized in vitro models allow two spatially distinct neural circuits or, more generally, two cell populations to develop functional connections and mimic key biological interactions.

Using Axion BioSystems' Maestro Pro and Edge systems, any scientist can now track functional network formation between neural co-cultures in the same well. Together, neural co-cultures on MEA plates create an ideal model for studying the functional connection between two networks, and the effects of synaptic blockade.

Tracking functional network formation between two neural populations
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A Culture-Insert 2 Well (ibidi, 80209) composed of silicone was used to culture two cortical neuron populations within the same well of a CytoView MEA 6-well plate (M384-tMEA-6W). At 7 days in vitro, the Culture-Insert 2 Well was removed, such that neurites could cross the cell-free gap and establish functional connections between the two distinct cortical networks. Experimental results from an optical image and activity map confirmed that a consistent, bi-directional functional connection between the two networks was formed within 10 days of Culture-Insert 2 Well removal.

Culture-Insert 2 Well in CytoView MEA plate for compartmentalized neural co-culture and functional connectivity research
Microscopy image showing neurite growth across a cell-free gap in an in vitro neural circuit and innervation model
MEA activity map showing synchronous activity between two spatially separated neural networks in vitro
MEA raster plot showing independent and synchronous network activity between two connected neural populations in vitro

A) Culture-Insert 2 Well (ibidi, 80209). B) Neurites project across the cell-free gap within 10 days of removing the insert. C) Activity map illustrating synchronous activity between the two spatially separated networks. D) Raster plot at 17 days in vitro illustrating independent network activity from Network 1 and Network 2, followed by a whole-well network event with Network 1 driving the activity in Network 2.

Model neuronal subtype interactions​
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Purpose: To demonstrate synaptic connectivity between different neuronal subtypes. Brain-on-a-chip models can shed light on the interaction between different brain regions.​

Brain-on-a-chip microscopy image showing neural cells cultured in compartmentalized MEA chambers for synaptic connectivity research
Fluorescence image of hippocampal and cortical neurons in a compartmentalized brain-on-a-chip MEA model
MEA raster plot showing early neural activity between cortical and hippocampal neuron chambers in a day 7 co-culture innervation assay
MEA raster plot showing increased synchronous neural firing between cortical and hippocampal neuron chambers after 21 days in vitro
MEA bar chart showing culture channel firing activity over time in cortical and hippocampal neuron co-culture chambers
MEA bar chart showing microchannel firing activity during synaptic connectivity formation in a neural co-culture model

Cortical and hippocampal neurons were cultured in separate chambers of the DuaLink MEA (Netri) and recorded on the Maestro MEA platform over 21 days.​

Results: Cultures increased synchronous firing between chambers as new synapses formed over time.​

Isolate and measure neurite activity​
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Purpose: To monitor growth and activity in neurites. Understanding the growth and function of neural projections could aid in the development neuroregenerative medicine.​

Fluorescence image showing neurite outgrowth across a compartmentalized MEA culture for neural innervation research
MEA bar chart showing increased culture channel activity as neurite projections develop in a neural innervation assay
MEA raster plot showing early neurite-associated neural activity in a compartmentalized cortical neuron culture at day 7
MEA raster plot showing increased neurite-associated neural activity in a compartmentalized cortical neuron culture at day 19

Cortical neurons were cultured in one chamber of the DuaLink MEA (Netri) and recorded on the Maestro MEA platform.​

Results: As new neurite projections covered the empty chamber, increasing activity was observed. This activity was synchronous to the activity in the cortical chamber.​

Assay Steps
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Neural co-culture assay workflow for using Culture-Insert 2 Well and CytoView MEA plates to monitor neural activity in vitro

Getting started with Maestro Pro and Edge couldn't be easier. Place the Culture-Insert 2 Well (ibidi, 80209) into the well of a CytoView MEA 6 well plate (M384-tMEA-6W). Culture your neurons in the desired compartments of the Culture-Insert 2 Well (Day 0). Remove the Culture-Insert 2 Well after the cells have attached to the surface of the MEA plate (~1-7 days). Load the MEA plate into the Maestro MEA system and allow the environmental chamber to automatically equilibrate. Analyze the neural activity in the MEA plate label-free and in real-time with AxIS Navigator Neural Module software (Day 2+).

Download Protocol

Download the Neural Activity Brochure for functional neural data and MEA-based neural circuit research applications