An operational concept for long-term cinemicrography of cells in mono- and co-culture under highly controlled conditions – The SlideObserver

• Published in: Journal of biotechnology 2012-05, Vol.159 (1-2), p.83-89
• Main Authors: Billecke, Nils, Raschzok, Nathanael, Rohn, Susanne, Morgul, Mehmet H., Schwartlander, Ruth, Mogl, Martina, Wollersheim, Sonja, Schmitt, Katharina R., Sauer, Igor M.
• Format: Article
• Language: English
• Subjects: Animals; Bioreactor; bioreactors; Cell Culture Techniques - methods; Cell Line, Tumor; Cell Proliferation - drug effects; Cells, Cultured; Cinemicrography; Coculture; Coculture Techniques - methods; Equipment Design; Hepatocytes; Hepatocytes - cytology; Hepatocytes - metabolism; Human Umbilical Vein Endothelial Cells; Humans; image analysis; in vitro studies; Intercellular Signaling Peptides and Proteins - pharmacology; Microscopy, Video - instrumentation; Microscopy, Video - methods; monitoring; Oxygen - metabolism; process control; Rats; Temperature; Time lapse
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2012.01.033

► Parallel time-lapse recording of different sets of cells in culture is challenging. ► We developed the SlideObserver for long-term culture and video-microscopy. ► The SlideObserver allows exact control of culture parameter. ► Seamless cinemicrography of primary cells for up to three weeks was possible. ► Long-term cinemicrography of cultured cells deliver insights into cellular behaviour. Cell morphology, proliferation and motility, as well as mono- and heterotypic cell-to-cell interactions, are of increasing interest for in vitro experiments. However, tightly controlling culture conditions whilst simultaneously monitoring the same set of cells is complicated. Moreover, video-microscopy of distinct cells or areas of cells over a prolonged period of time represents a technical challenge. The SlideObserver was designed for cinemicrography of cells in co-and monoculture. The core elements of the system are the SlideReactors, miniaturised hollow fibre-based bioreactors operated in closed perfusion loops. Within the SlideReactors, cells can be cultured under adaptable conditions as well as in direct- and indirect co-culture. The independent perfusion loops enable controlled variation of parameters such as medium, pH, and oxygenation. A combined automated microscope stage and camera set-up allows for micrograph acquisition of multiple user-defined regions of interest within the bioreactor units. For proof of concept, primary cells (HUVEC, human hepatocytes) and cell lines (HuH7, THP-1) were cultured under stable and varying culture conditions, as well as in mono- and co-culture. The operational system enabled non-stop imaging and automated control of process parameters as well as elective manipulation of either reactor. As opposed to non-perfused culture systems or comparable devices for cinemicrographic analysis, the SlideObserver allows simultaneous morphological monitoring of an entire culture of cells in multiple bioreactors.