caa a22 4500 605504911 CHVBK 20210128100619.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6855-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6855-z Lactate and glucose concomitant consumption as a self-regulated pH detoxification mechanism in HEK293 cell cultures [Elektronische Daten] [Leticia Liste-Calleja, Martí Lecina, Jonatan Lopez-Repullo, Joan Albiol, Carles Solà, Jordi Cairó] One of the most important limitations of mammalian cell-based processes is the secretion and accumulation of lactate as a by-product of their metabolism. Among the cell lines commonly used in industrial bioprocesses, HEK293 has been gaining importance over the last years. Up recently, HEK293 cells were known to consume lactate in late stages of cell culture usually when glucose and/or glutamine were depleted from media. Remarkably, in both scenarios, no significant cell growth was reported. However, we have observed a different metabolic behavior regarding lactate production and consumption in HEK293 cultures. HEK293 cells were able to co-metabolize glucose and lactate simultaneously, even in exponentially growing cell cultures. Our deep study of the effects of environmental conditions on lactate metabolism revealed that pH was the key to trigger the metabolic shift from lactate production to lactate and glucose concomitant consumption. Remarkably, this shift could be triggered at will when pH was set at 6.8. Even more interesting was the fact that lowering pH to 6.6 and supplementing media with exogenous lactate resulted in co-consumption of glucose and lactate from the beginning of cell culture, without affecting cell growth or protein productivity. On the contrary, cell growth was clearly hampered at this low pH if extracellular lactate was lacking. From our results, we hypothesize that HEK293 cells metabolize extracellular lactate as a strategy for pH detoxification, by means of co-transporting extracellular protons together with lactate into the cytosol. This novel hypothesis for unraveling lactate metabolism in HEK293 cells could open a door to re-direct genetic engineering strategies in order to obtain more efficient cell lines and also to further develop animal cell technology applications. Springer-Verlag Berlin Heidelberg, 2015 HEK293 nationallicence Lactate nationallicence Metabolic shift nationallicence pH nationallicence Warburg effect nationallicence Co-metabolism nationallicence Liste-Calleja Leticia Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Lecina Martí Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Lopez-Repullo Jonatan Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Albiol Joan Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Solà Carles Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Cairó Jordi Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 9951-9960 0175-7598 99:23<9951 2015 99 253 https://doi.org/10.1007/s00253-015-6855-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6855-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liste-Calleja Leticia Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 700 1- Lecina Martí Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 700 1- Lopez-Repullo Jonatan Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 700 1- Albiol Joan Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 700 1- Solà Carles Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 700 1- Cairó Jordi Chemical Engineering Department, Universitat Autònoma de Barcelona, Edifici Q, Campus Bellatera. Carretera de Cerdanyola, s/n. 08193 Bellatera, Barcelona, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 9951-9960 0175-7598 99:23<9951 2015 99 253