Overflow Microfluidic Networks for Open and Closed Cell Cultures on Chip

摘要

Microfluidics have a huge potential in biomedical re-nsearch, in particular for studying interactions among cellnpopulations that are involved in complex diseases. Here,nwe present “overflow” microfluidic networks (oMFNs) forndepositing, culturing, and studying cell populations, whichnare plated in a few microliters of cell suspensions in onenor several open cell chambers inside the chip and sub-nsequently cultured for several days in vitro (DIV). Afternthe cells have developed their phenotype, the oMFN isnclosed with a lid bearing microfluidic connections. Thensalient features of the chips are (1) overflow zones aroundnthe cell chambers for drawing excess liquid by capillaritynfrom the chamber during sealing the oMFN with the lid,n(2) flow paths from peripheral pumps to cell chambersnand between cell chambers for interactive flow control,n(3) transparent cell chambers coated with cell adhesionnmolecules, and (4) the possibility to remove the lid fornstaining and visualizing the cells after, for example,nfixation. Here, we use a two-chamber oMFN to show thenactivation of purinergic receptors in microglia grown innone chamber, upon release of adenosine triphosphaten(ATP) from astrocytes that are grown in another chambernand challenged with glutamate. These data validate oM-nFNs as being particularly relevant for studying primaryncells and dissecting the specific intercellular pathwaysninvolved in neurodegenerative and neuroinflammatorynbrain diseases.