posted on 2023-12-27, 19:36authored byLydia Moll, Johan Pihl, Mattias Karlsson, Paul Karila, Camilla I. Svensson
Compartmentalized cell cultures (CCCs) provide the possibility
to study mechanisms of neurodegenerative diseases, such as spreading
of misfolded proteins in Alzheimer’s or Parkinson’s
disease or functional changes in, e.g., chronic pain, in vitro. However,
many CCC devices do not provide the necessary capacity for identifying
novel mechanisms, targets, or drugs in a drug discovery context. Here,
we present a high-capacity cell culture microtiter microfluidic plate
compliant with American National Standard Institute of the Society
for Laboratory Automation and Screening (ANSI/SLAS) standards that
allows to parallelize up to 96 CCCs/experimental units, where each
experimental unit comprises three microchannel-connected compartments.
The plate design allows the specific treatment of cells in individual
compartments through the application of a fluidic barrier. Moreover,
the compatibility of the plate with neuronal cultures was confirmed
with rodent primary as well as human-induced pluripotent stem cell-derived
neurons of the central or peripheral nervous system for up to 14 days
in culture. Using immunocytochemistry, we demonstrated that the plate
design restricts neuronal soma to individual compartments, while axons,
but not dendrites, can grow through the connecting microchannels to
neighboring compartments. In addition, we show that neurons are spontaneously
active and, as deemed by the appearance of synchronous depolarizations
in neighboring compartments, are synaptically coupled. In summary,
the design of the microfluidic plate allows for both morphological
and functional studies of neurological in vitro cultures with increased
capacity to support identification of novel mechanisms, targets, or
drugs.