Compartmentalized Primary Murine Neuron Culture Using Plastic Microfluidic Chips
Transcription
Take a plastic microfluidic chip comprising somatic reservoirs and compartments, as well as axonal reservoirs and compartments, connected by microgrooves.
Add a pre-coating solution to saturate the compartments and grooves, preventing air bubble entrapment.
Aspirate the solution, leaving the main compartments filled. Wash with buffer.
Coat the wells with a synthetic polymer that facilitates neuronal adhesion. Wash with buffer and add culture media to prevent the chip from drying.
To seed a suspension of rat hippocampal neurons, aspirate the media.
Load the suspension into the somatic reservoirs. The neurons enter and attach to the somatic compartment.
Add neuronal culture media to the wells and incubate. Growth factors present in the media facilitate neuronal growth.
Gradually, the neurons extend their axons, which grow into the axonal compartment through the microgrooves.
The narrowness of the grooves prevents the entry of neuronal cell bodies, ensuring the physical separation of somatic and axonal regions.
