Host-pathogen interactions represent a signicant areao biomedical research, encompassing the study o howviruses, bacteria, and other organisms cause inectionsand diseases or alter normal physiology o host cellsand tissues
. As the eld o cellular systems biologyadvances, there is increasing interest in using
cellculture models to study host-pathogen interactions
.The proper study o host-pathogen interactions requirescareul control o the cell environment to simulatephysiologic conditions. An
model that canreplicate inection parameters, including fow rate,exposure time, solution type, and timed introductiono therapeutic agents while sustaining long-term livecell microscopy enables experiments that are dicultor impossible with standard methods. In many cases,it is crucial to precisely manipulate the exposureo pathogen to the cultured host cells: too low anexposure may prevent appropriate interactions, whiletoo high an exposure could cause conounding eects(such as bacterial overgrowth and nutrient starvation)independent o the desired host-pathogen interaction.In addition, inormation on the eect o exposuretimes and fow conditions could prove valuable orunderstanding the mechanisms o disease or manyinectious agents. A urther challenge in host-pathogenresearch is the ability to sustain inections
, sincecultured host cells oten de-dierentiate in culture andno longer support pathogen responses ound
. Forcertain pathogens, the time scale o inection even underoptimal conditions could take days or weeks, placingadditional burdens on the cell culture method. Recentadvances in microfuidics-based cell culture technologiesoer potential solutions to these problems
.The CellASIC™ ONIX Microfuidic System is well suitedor host-pathogen studies by providing a stable, long-term culture environment or host cells (includingprimary cells) with controlled pathogen exposure. Thecontinuous perusion ormat and the ability to switchmedia solutions enable wash-out o pathogens romthe chamber and subsequent monitoring o host cellresponse over many days. The enclosed small volume o the culture chamber also provides practical advantagesor working with inectious agents during live cellimaging. The design o the microfuidic plates enablesthe dynamics o inection to be tracked using live cellimaging on standard inverted microscopes.Here, we demonstrate a host-pathogen experimentusing human intestinal cells inected with engineered
strains (Figure 1). Both an invasive and non-invasive bacterial strain were monitored or long-terminection with time-lapsed imaging up to 24 hours inthe CellASIC™ M04S microfuidic plate.
Long-term, live cell imaging o host-pathogen interactions usingthe CellASIC
Human colonadenocarcinoma cells(HT-29) cultured inthe M04S microuidicchamber and inectedwith an invasive strain o
(A) Phase contrastimage showing apoptoticcells, and (B) uorescenceimage showing invadedbacteria (red) and cellviability (green). Imageswere acquired with a100X objective lens.
EMD Millipore is a division o Merck KGaA, Darmstadt, Germany