The reproducibility of high-throughput cell-based assays is dependent on having a consistent source of cells for each experiment. Developing an understanding of the nature of cells growing in vitro and factors that influence their responsiveness to test compounds will contribute to the development of reproducible cell-based assays. Using good cell culture practices and establishing standard operating procedures (SOPs) for handling cultures can eliminate several potential contributors to variability in the responsiveness and performance of cells. The SOPs for handling each cell type must have clear and detailed instructions that can be understood and followed among different laboratories. The SOPs should include documenting the source of cells and authenticating their identity, both of which have become required to achieve peer acceptance of experimental data. Variability caused by biological issues such as phenotypic drift can be reduced by using standardized subculture procedures or using cryopreserved cells to set up experiments. Variability caused by inconsistent dispensing of cells per well and edge effects can be identified by measuring how many cells are present and whether they are alive or dead. Multiplex methods for real-time measurement of viable or dead cell number in each sample can be used for normalizing data and determining if proliferation or cytotoxicity has occurred during the experiment. Following good cell culture practices will go a long way toward executing reproducible cell-based assays. Resources will be included describing good cell culture practices, cell line authentication, and multiplex determination of cell number as an internal control.
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http://dx.doi.org/10.1177/24725552211039754 | DOI Listing |
Mar Drugs
January 2025
Institut de Recherche pour le Développement (IRD), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), P.O. Box 6570, 98702 Faa'a, Tahiti, French Polynesia.
Ciguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus . Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrices, are numerous in the literature. However, little information on their effectiveness for nonpolar CTXs is available, yet these congeners, contributing to the risk of CP, are required for the establishment of effective food safety monitoring programs.
View Article and Find Full Text PDFCurr Issues Mol Biol
December 2024
Department of Clinical Sciences and Translational Medicine, University of Rome 'Tor Vergata', Via Montpellier 1, 00133 Rome, Italy.
Cancer cells demonstrate remarkable resilience by adapting to oxidative stress and undergoing metabolic reprogramming, making oxidative stress a critical target for cancer therapy. This study explores, for the first time, the redox-dependent anticancer effects of Polydatin (PD), a glucoside derivative of resveratrol, on the human Osteosarcoma (OS) cells SAOS-2 and U2OS. Using cell-based biochemical assays, we found that cytotoxic doses of PD (100-200 µM) promote ROS production, deplete glutathione (GSH), and elevate levels of both total iron and intracellular malondialdehyde (MDA), which are key markers of ferroptosis.
View Article and Find Full Text PDFDent J (Basel)
January 2025
Institute of Dentistry, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
: Halitosis is primarily caused by the activity of oral microorganisms. In this study, we employed metagenomic sequencing and metabolomic approaches to investigate the differences in salivary microbiota and metabolite profiles between individuals with halitosis and periodontitis and healthy controls. Additionally, we expanded the study to examine how oral malodorous compounds interact with human oral squamous carcinoma (HSC-4) cells.
View Article and Find Full Text PDFCells
January 2025
Department of Experimental Medicine, Sapienza University of Rome, 00161 Rome, Italy.
Adipose-derived mesenchymal stem cells (ASCs) are commonly employed in clinical treatment for various diseases due to their ability to differentiate into multi-lineage and anti-inflammatory/immunomodulatory properties. Preclinical studies support their use for bone regeneration, healing, and the improvement of functional outcomes. However, a deeper understanding of the molecular mechanisms underlying ASC biology is crucial to identifying key regulatory pathways that influence differentiation and enhance regenerative potential.
View Article and Find Full Text PDFFront Pharmacol
January 2025
Bioactive Botanical Research Laboratory, Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, RI, United States.
Oleuropein is a phenolic compound commonly found in cosmetic ingredients including olive leaves and jasmine flowers with various skin-beneficial effects. Here, we evaluated oleuropein's anti-inflammatory and antioxidant activities in human skin cells. In a cell-based inflammasome model with human monocytes (THP-1 cells), oleuropein (12-200 µM) reduced proinflammatory cytokine interleukin (IL)-6 by 38.
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