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Caption : supplementary figure 1 a) schematic diagram of the three most relevant metabolic pathways to maintain cell homeostasis. during short-term 7-minute (b), and long-term 90-minute periods (c), the perfusion rate was altered and time-resolved metabolic levels were recorded. while glycolytic metabolites accumulate or are degraded after a short time, for glutamine and free fatty acids a change can only be detected after a long-term change of the perfusion rate. y-axis represents relative metabolite content normalized to their initial values. experiments with same observations as shown in b and c were measured in two independent replicates each.

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Caption : supplementary table 1 measuring lactate/glucose ratio helps to detect hypoxic cell culture condition. in the first line, 20 million cells were incubated conventionally in a petri dish inside the incubator. in lines 2 to 6, the glucose and lactate values at the outlet of the perfused medium were measured in two biological replicas for each different perfusion rate. glucose and lactate values were determined using cobas® b 123 at outlet of the perfused bioreactor system. reference activity of 3 million cells (80 % confluence) cultured under proliferative 2d conditions are shown in the last line of the table. in order to increase the comparability in the table, all shown results were normalized to the corresponding activity of 1 million fibroblasts.