An operational concept for long-term cinemicrography of cells in mono- and co-culture under highly controlled conditions--the SlideObserver.
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.
Department of General, Visceral, and Transplantation Surgery, Charité-Campus Virchow, Universitätsmedizin Berlin, Virchow, Augustenburgerplatz 1, Berlin, Germany. nils.billecke@charite.de
Nils
N
Billecke
Nathanael
N
Raschzok
Susanne
S
Rohn
Mehmet H
MH
Morgul
Ruth
R
Schwartlander
Martina
M
Mogl
Sonja
S
Wollersheim
Katharina R
KR
Schmitt
Igor M
IM
Sauer
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D004867:Equipment Design; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D018715:Microscopy, Video / Q000295:instrumentation* / Q000379:methods; D010100:Oxygen / Q000378:metabolism; D051381:Rats; D013696:Temperature
D036341:Intercellular Signaling Peptides and Proteins; D010100:Oxygen