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

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• 作者：Nils Billecke^a ; ^{nils.billecke@charite.de} ; Nathanael Raschzok^a ; ^{nathanael.raschzok@charite.de} ; Susanne Rohn^a ; ^{susanne.rohn@charite.de} ; Mehmet H. Morgul^b ; ^{haluk.morgul@uniklinik-leipzig.de} ; Ruth Schwartlander^c ; ^{ruth.schwartlaender@mat.ethz.ch} ; Martina Mogl^a ; ^{martina.mogl@charite.de} ; Sonja Wollersheim^d ; ^{sonja.wollersheim@charite.de} ; Katharina R. Schmitt^d ; ¹ ; ^{katharina.schmitt@charite.de} ; Igor M. Sauer^a ; ¹ ; ^{igor.sauer@charite.de}
• 关键词：Time lapse ; Cinemicrography ; Hepatocytes ; Coculture ; Bioreactor
• 刊名：Journal of Biotechnology
• 出版年：2012
• 期刊代码：53_01681656
• 类别：imm
• 出版时间：31 May, 2012
• 卷：159
• 期：1-2
• 页码：83-89
• 文件大小：1922 K

摘要

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.