High throughput, non-invasive and dynamic toxicity screening on adherent cells using respiratory measurements

In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor

“…Oxygen uptake rate measurement in well‐mixed mammalian cell bioreactors was previously reported using liquid or gas phase balances (Eyer et al , 1995 Oeggerli et al , 1995) and was applied for culture medium optimization purposes (Deshpande et al , 2004). Moreover, the online monitoring of oxygen was also shown to be a promising tool in terms of toxicity testing in hepatocyte cultures (Beckers et al , 2010; Niklas et al , 2009; Noor et al , 2009). In a perfused multiwell plate designed for 3D cultivation on scaffolds, oxygen consumption and oxygen transport was modelled and consumption rates were measured using optical probes, showing the possibility of long‐term maintenance of liver cells on the basis of viability and immunostaining for albumin (Domansky et al , 2010).…”

Incipient cytotoxicity: A time-independent measure of cytotoxic potency in vitro

“…Exposure durations in in vitro toxicity assays are more or less arbitrarily chosen and range from one hour or less to several days (see, for instance, the international MEIC project, Clemedson et al, 1996 or the ToxCast project, Judson et al, 2010). Various studies with different chemicals and in vitro systems, however, revealed that the concentration of a chemical found to be toxic can largely depend on the exposure time used (Riddell et al, 1986;Seibert et al, 2004;Gülden et al, 2005;Komissarova et al, 2005;Fotakis and Timbrell, 2006;Beckers et al, 2010;Gülden et al, 2010;, Inácio et al, 2011 Fotakis andTimbrell, 2006;Beckers et al, 2010;Gülden et al, 2010;Inácio et al, 2011). This calls into question the relevance of in vitro toxic potency measures after arbitrary times and their suitability for the above mentioned purposes.…”

Respirometry: Correcting for Diffusion and Validating the Use of Plastic Multiwell Plates with Integrated Optodes

“…In recent years, the use of integrated luminescencebased optochemical oxygen (Oj) sensors (optodes) to measure P02 has gained favor over formerly used Clarke-type electrochemical sensors in respirometric studies because the former do not consume O2, require low maintenance, and provide more stable measurements. In closed-system designs, the simultaneous measurement of replicates using polystyrene multiwell plates as well as glass vials with integrated optodes is increasingly used to measure R in small animals and cells (Deshpande and Heinzle 2004;Köster et al 2008;Abaci et al 2010;Beckers et al 2010;White et al 2011). Ideally, respirometry chambers should be nondiffusible for gases.…”