To overcome the problems of endpoint tests routinely required for EC50 determination, we utilized a novel automated high-content workstation and calculated a time-resolved EC50 value of MCF-7 mamma carcinoma cells treated with a pharmacologic agent. Measuring parameters were the cellular oxygen consumption and the extracellular acidification. These parameters were detected in real-time and label free with optochemical sensor spots in a modified 24-well sensor plate. In particular, the objective was to compare the measuring data of the workstation with a classical standard resazurin cell assay and to transfer the benefit of continuously recorded metabolic data of the workstation to practical time-resolved information about the effect of the applied active reagent (doxorubicin). MCF-7 cells were treated with a broad range of doxorubicin concentrations (100 μM to 1 nM) over 24 h and cellular activities were investigated with both, the resazurin assay and the automated workstation. Twenty-four hours after treatment, the resazurin assay showed an EC50 value (6.3 μM) which was about one decade above the value obtained from oxygen consumption rate (0.37 μM) and extracellular acidification rate (0.72 μM), measured with the workstation. Presumably, the differences are due to the different metabolic nature and regulation behind these measuring parameters. By polynomial fitting of continuously recorded metabolic data, we were able to point out a dynamic, time-resolved EC50 characteristic for different time points. The workstation is a powerful tool to record in vitro kinetic data of pharmacologic effects in vital cells in an automated experimental run.
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http://dx.doi.org/10.1016/j.tiv.2011.05.031 | DOI Listing |
Biofilm
December 2024
Department of Biomaterials, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Biodegradable polymeric coatings are being explored as a preventive strategy for orthopaedic device-related infection. In this study, titanium surfaces (Ti) were coated with poly-D,L-lactide (PDLLA, (P)), polyethylene-glycol poly-D,L-lactide PEGylated-PDLLA, (PP20)), or multi-layered PEGylated-PDLLA (M), with or without 1 % silver sulfadiazine. The aim was to evaluate their cytocompatibility, resistance to biofilm formation, and their potential to enhance the susceptibility of any biofilm formed to antibiotics.
View Article and Find Full Text PDFHeliyon
December 2024
Human and Animal Physiology, Department Animal Sciences, Wageningen University, De Elst 1, 6708WD, Wageningen, the Netherlands.
Label-free imaging is routinely used during cell culture because of its minimal interference with intracellular biology and capability of observing cells over time. However, label-free image analysis is challenging due to the low contrast between foreground signals and background. So far various deep learning tools have been developed for label-free image analysis and their performance depends on the quality of training data.
View Article and Find Full Text PDFBMC Cancer
December 2024
CeMOS, Mannheim University of Applied Sciences, 68163, Mannheim, Germany.
Background: The growth and drug response of tumors are influenced by their stromal composition, both in vivo and 3D-cell culture models. Cell-type inherent features as well as mutual relationships between the different cell types in a tumor might affect drug susceptibility of the tumor as a whole and/or of its cell populations. However, a lack of single-cell procedures with sufficient detail has hampered the automated observation of cell-type-specific effects in three-dimensional stroma-tumor cell co-cultures.
View Article and Find Full Text PDFSci Total Environ
January 2025
Institute for Environmental Assessment and Water Research (IDAEA-CSIC), Jordi Girona, 18, 08034 Barcelona, Spain. Electronic address:
The acoustic startle response (ASR) is leaded by a sudden and intense acoustic stimulus. ASR has several forms of plasticity, including habituation and sensorimotor gating. Although ASR and its plasticity have been intensively studied in zebrafish (Danio rerio) larvae, information in adult zebrafish is still very scarce.
View Article and Find Full Text PDFPLoS Biol
December 2024
School of Biosciences, University of Sheffield, Western Bank, Sheffield, United Kingdom.
The interaction between cancer cells and the extracellular matrix (ECM) plays a pivotal role in tumour progression. While the extracellular degradation of ECM proteins has been well characterised, ECM endocytosis and its impact on cancer cell progression, migration, and metastasis is poorly understood. ECM internalisation is increased in invasive breast cancer cells, suggesting it may support invasiveness.
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