Proliferation characteristics of cells cultured under periodic versus static conditions
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In vitro culture models have become an indispensable tool for assessing a vast variety of biological questions in many scientific fields. However, common in vitro cultures are maintained under static conditions, which do not reflect the in vivo situation and create a non-physiological environment. To assess whether the growth characteristics of cells cultured at pulsed-perfused versus static conditions differ, we observed the growth of differentially cultured cells in vitro by life-cell time-lapse imaging of recombinant HEK293YFPI152L cells, stably expressing yellow fluorescent protein. Cells were grown for ~ 30 h at 37 °C and ambient CO2 concentration in biochips mounted into a custom-designed 3D printed carrier and were imaged at a rate of ten images per hour using a fluorescence microscope with environment control infrastructure. Cells in one chip were maintained under static conditions whereas cells in another chip were recurrently perfused with fresh media. Generated image series were quantitatively analyzed using a custom-modified cell detection software. Imaging data averaged from four biological replicates per culturing condition demonstrate that cells cultured under conventional conditions exhibit an exponential growth rate. In contrast, cells cultured in periodic mode exhibited a non-exponential growth rate. Our data clearly indicate differential growth characteristics of cells cultured under periodic versus static conditions highlighting the impact of the culture conditions on the physiology of cells in vitro.
KeywordsCell growth Biochip Microfluidics HEK293 YFPI152L Long-term time-lapse microscopy
The authors gratefully acknowledge funding of the Staedtler Stiftung and ongoing support from the Erlangen Graduate School in Advanced Optical Technologies (SAOT) by the German Research Foundation (DFG) in the framework of the German Excellence Initiative. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
J.W. and D.F.G. conceived the study. S.A.M. and D.F.G. conducted imaging experiments. D.F.G. analyzed and displayed imaging data. D.F.G. and J.W. and wrote the paper. All authors commented and agreed on the manuscript.
Compliance with ethical standards
Conflict of interest
JW is CEO and shareholder of Cellasys GmbH.
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