Assaying Proliferation Characteristics of Cells Cultured Under Static Versus Periodic Conditions
Gilbert D, Friedrich O, Wiest J (2023)
Publication Type: Book chapter / Article in edited volumes
Publication year: 2023
Journal
Publisher: Humana Press Inc.
Edited Volumes: Cell Viability Assays (2nd Edt.)
Series: Methods in Molecular Biology
Book Volume: 2644
Pages Range: 35-45
DOI: 10.1007/978-1-0716-3052-5_3
Abstract
Two-dimensional in vitro culture models are widely being employed for assessing a vast variety of biological questions in different scientific fields. Common in vitro culture models are typically maintained under static conditions, where the surrounding culture medium is replaced every few days—typically every 48 to 72 h—with the aim to remove metabolites and to replenish nutrients. Although this approach is sufficient for supporting cellular survival and proliferation, static culture conditions do mostly not reflect the in vivo situation where cells are continuously being perfused by extracellular fluid, and thus, create a less-physiological environment. In order to evaluate whether the proliferation characteristics of cells in 2D culture maintained under static conditions differ from cells kept in a dynamic environment, in this chapter, we provide a protocol for differential analysis of cellular growth under static versus pulsed-perfused conditions, mimicking continuous replacement of extracellular fluid in the physiological environment. The protocol involves long-term life-cell high-content time-lapse imaging of fluorescent cells at 37 °C and ambient CO2 concentration using multi-parametric biochips applicable for microphysiological analysis of cellular vitality. We provide instructions and useful information for (i) the culturing of cells in biochips, (ii) setup of cell-laden biochips for culturing cells under static and pulsed-perfused conditions, (iii) long-term life-cell high-content time-lapse imaging of fluorescent cells in biochips, and (iv) quantification of cellular proliferation from image series generated from imaging of differentially cultured cells.
Authors with CRIS profile
Daniel Gilbert
Lehrstuhl für Medizinische Biotechnologie
Oliver Friedrich
Lehrstuhl für Medizinische Biotechnologie
Involved external institutions
Germany (DE)How to cite
APA:
Gilbert, D., Friedrich, O., & Wiest, J. (2023). Assaying Proliferation Characteristics of Cells Cultured Under Static Versus Periodic Conditions. In Friedrich O, Gilbert DF (Eds.), Cell Viability Assays (2nd Edt.). (pp. 35-45). Humana Press Inc..
MLA:
Gilbert, Daniel, Oliver Friedrich, and Joachim Wiest. "Assaying Proliferation Characteristics of Cells Cultured Under Static Versus Periodic Conditions." Cell Viability Assays (2nd Edt.). Ed. Friedrich O, Gilbert DF, Humana Press Inc., 2023. 35-45.
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