Abstract
In vitro neuronal models are essential for studying neurological physiology, disease mechanisms and potential treatments. Most in vitro models lack controlled vasculature, despite its necessity in brain physiology and disease. Organ-on-chip models offer microfluidic culture systems with dedicated micro-compartments for neurons and vascular cells. Such multi-cell type organs-on-chips can emulate neurovascular unit (NVU) physiology, however there is a lack of systematic data on how individual cell types are affected by culturing on microfluidic systems versus conventional culture plates. This information can provide perspective on initial findings of studies using organs-on-chip models, and further optimizes these models in terms of cellular maturity and neurovascular physiology. Here, we analysed the transcriptomic profiles of co-cultures of human induced pluripotent stem cell (hiPSC)-derived neurons and rat astrocytes, as well as one-day monocultures of human endothelial cells, cultured on microfluidic chips. For each cell type, large gene expression changes were observed when cultured on microfluidic chips compared to conventional culture plates. Endothelial cells showed decreased cell division, neurons and astrocytes exhibited increased cell adhesion, and neurons showed increased maturity when cultured on a microfluidic chip. Our results demonstrate that culturing NVU cell types on microfluidic chips changes their gene expression profiles, presumably due to distinct surface-to-volume ratios and substrate materials. These findings inform further NVU organ-on-chip model optimization and support their future application in disease studies and drug testing.
Highlights
In vitro neuronal models are essential for studying neurological physiology, disease mechanisms and potential treatments
We demonstrate that the implemented culture system affects gene expression in a cell type-specific manner, showing decreased cell division in endothelial cells, increased cell adhesion in iNeurons and astrocytes, and increased maturity of iNeurons when cultured on a microfluidic chip
As we directly compare cells cultured on the microfluidic chips and cells cultured in wells, we only evaluated iNeuron and astrocyte co-cultures and Human umbilical vein endothelial cells (HUVECs) monocultures in this microfluidic chip
Summary
In vitro neuronal models are essential for studying neurological physiology, disease mechanisms and potential treatments. Current in vitro neuronal models are typically based on a two-dimensional (2D) cell layer cultured in a microwell plate, which are not designed to controllably mimic the three-dimensional (3D) geometry of nervous tissue They lack a vascular compartment which prevents their use in studies of neurovascular disease or blood–brain. Though useful in studying cell–cell interactions, such transwell systems lack key aspects of the geometrical and physical microenvironment of brain tissue They lack blood vessels with perfusable lumens, and the cell-to-volume ratio of the cell culture medium is non-physiological[7]. Brain organoids are a more complex in vitro neuronal models, consisting of multicellular tissues with a complex 3D geometry than can include vascular structures[8,9,10] These models are very useful when studying cell–cell interactions and the CNS microenvironment, difficulties in controlling their formation results in high variability. Using patient-specific human induced pluripotent stem cell (hiPSC)-derived cells, these devices can considerably aid the development of personalized m edicine[12,15,16,17,18,19]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
