Once recovered, spheroids exhibit a metabolism that is stable from 18 to at least 42 days in culture and produce urea, cholesterol and ATP at physiological levels (i.e. Previously, we have shown that HepG2/C3A cells need 18 days to complete the transition from exponential growth to dynamic equilibrium. During culture, these clusters round up and are therefore termed 3D microgravity ‘spheroids’ in this manuscript. After incubation these clusters are released into the rotating culture chamber and then treated in essentially the same way as one would treat cells in a culture flask. The cell clusters are initiated by centrifuging cells into inverted pyramidal indentations in a special culture plate. In this system a culture chamber is rotated on a clinostat in a standard cell culture incubator (also known as microgravity cultures). We have used a cell culture system based on the maintenance of cell clusters in suspension to induce cells to reach the dynamic equilibrium. The second is characterized by cells which have reached a dynamic equilibrium, growing either in 3D tissues or tissue-like conglomerates, in which the doubling time is long (a month or longer) and in which trypsin is rarely, if ever, used. The first is characterized by exponential growth of cultures using classical 2D culture techniques typically involving a cycle of trypsinisation, inoculation, growth and re-trypsinisation and a short doubling time (usually a few days). There are two extreme conditions for cell growth. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. ![]() All relevant data are within the paper and its Supporting Information files.įunding: University of Southern Denmark. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: The authors confirm that all data underlying the findings are fully available without restriction. Received: ApAccepted: AugPublished: September 15, 2014Ĭopyright: © 2014 Wrzesinski et al. PLoS ONE 9(9):Įditor: Manlio Vinciguerra, University College London, United Kingdom (2014) The Cultural Divide: Exponential Growth in Classical 2D and Metabolic Equilibrium in 3D Environments. Citation: Wrzesinski K, Rogowska-Wrzesinska A, Kanlaya R, Borkowski K, Schwämmle V, Dai J, et al.
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