Preparation of neuronal co-cultures with single cell precision
Journal
Journal of Visualized Experiments
Journal Issue
87
Date Issued
2014-05-20
Author(s)
Abstract
Microfluidic embodiments of the Campenot chamber have attracted great interest from the neuroscience community. These interconnected co-culture platforms can be used to investigate a variety of questions, spanning developmental and functional neurobiology to infection and disease propagation. However, conventional systems require significant cellular inputs (many thousands per compartment), inadequate for studying low abundance cells, such as primary dopaminergic substantia nigra, spiral ganglia, and Drosophilia melanogaster neurons, and impractical for high throughput experimentation. The dense cultures are also highly locally entangled, with few outgrowths (<10%) interconnecting the two cultures. In this paper straightforward microfluidic and patterning protocols are described which address these challenges: (i) a microfluidic single neuron arraying method, and (ii) a water masking method for plasma patterning biomaterial coatings to register neurons and promote outgrowth between compartments. Minimalistic neuronal co-cultures were prepared with high-level (>85%) intercompartment connectivity and can be used for high throughput neurobiology experiments with single cell precision.
Subjects
Alzheimer and parkinson diseases | Biomaterial patterning | Co-culture | Compartmentalization | High throughput screening | Issue 87 | Microfluidic arraying | Neurite outgrowth | Neuroscience | Single cell
Type
journal article