Compartmentalization of Human Stem Cell-Derived Neurons within Pre-Assembled Plastic Microfluidic Chips

Area of Science:

• Neuroscience
• Stem Cell Biology
• Microfluidics

Background:

• Microfluidic devices are standard for compartmentalizing cultured neurons in neuroscience.
• Human stem cell-derived neurons offer a powerful model for studying neuronal development and function.

Purpose of the Study:

• To provide a protocol for using pre-assembled cyclic olefin copolymer (COC) multi-compartment chips for culturing human stem cell-derived neurons.
• To demonstrate the compatibility of these chips with standard microscopy and experimental procedures.

Main Methods:

• Neurons were differentiated from human neural stem cells (NSCs) into glutamatergic neurons within COC chips.
• Chips were maintained for 5 weeks to allow for neuronal development, including synapse and dendritic spine formation.
• Experimental procedures such as viral labeling, microenvironment manipulation, axotomy, and immunocytochemistry were performed on the cultured neurons.

Main Results:

• Successful differentiation and long-term maintenance of human stem cell-derived glutamatergic neurons in COC microfluidic chips.
• Demonstrated development of synapses and dendritic spines, indicating functional neuronal maturation.
• Validated the utility of the chips for various experimental manipulations common in neuroscience.

Conclusions:

• COC microfluidic chips provide a robust and versatile platform for culturing and experimenting with human stem cell-derived neurons.
• This protocol facilitates advanced neuroscience research by enabling detailed study of neuronal development and function in a controlled microenvironment.