ISBN 9783843936859

978-3-8439-3685-9, Reihe Ingenieurwissenschaften

Frank Bunge
On-chip Mammalian Cell Cultivation and Monitoring

155 Seiten, Dissertation Universität Bremen (2018), Softcover, B5

Zusammenfassung / Abstract

The biological and medical progress is, among other factors, also enabled by powerful analysis devices. In the future, new tools will be based on microfluidics to assay mammalian cells. In this thesis, novel concepts for long-term cultivation and monitoring of cells are investigated in the context of Lab-on-a-Chip, which are compact analysis systems in which all relevant processes are integrated. In general, the focus of this work is laid on entire systems instead of single components or functionalities. In particular, the requirements for the presented systems are the usage of non-cytotoxic materials, the fabrication based on clean room processes, the integration of functionalities to avoid bulky and expensive external equipment wherever possible and easy-to-use interfaces. The cell monitoring is limited to the measurement of oxygen uptake of mammalian cells which indicates the metabolism, being a key process of living cells.

In this thesis, two concepts are discussed for the on-chip cultivation of adherent mammalian cells. Both concepts contain porous membranes to supply the cells with nutrients and gases by diffusion. The horizontal concept is based on diffusion processes in horizontal direction through membranes made of agarose hydrogel. In contrast, the supply takes place in vertical direction through a membrane of anodised aluminium oxide in the vertical concept. The successful on-chip passaging is shown with MDCK- and HaCaT cells over 48 h and 14 days, respectively.

The oxygen consumption rate is determined by measuring the decay of the oxygen concentration inside a closed microfluidic chip. This sensing is based on an oxygen sensitive, phosphorescent dye that is embedded in a thin, porous film. In addition, electrical heating elements are integrated into the chip while all optical components are part of the assembly. With this system, the oxygen consumption rate of HaCaT cells is determined for different temperatures.

The discussed concepts and results show that microfluidic devices are well suited for on-chip cell cultivation and monitoring despite their requirements.