On-line oxygen uptake rate and culture viability measurement of animal cell culture using microplates with integrated oxygen sensors

摘要

O_2 uptake rates of animal cells (Chinese hamster ovary-CHO) were measured in 96-well microtiter plates by integrating with fluorescent sensors thereby measuring fluorescence intensity ratios of an O_-sensitive and an insensitive fluorophor.O_2 consumption rate was estimated from measured dissolved O_2 and from O_2 mass transfer coefficient determined in advance.Specific uptake decreased with time from 3.2X10~(-13) mol O_2 cell~(_1) h~(-1) at 15 h cultivation to 1.8X10~(-13) mol O_2 cell~(-1) h~(-1) at 48 h.Specific O_2 uptake was also determined by sampling from a spinner-flask culture giving identical values.A cell viability assay for cultures based on O_2 measurements is described in which cells are incubated outside the fluorescence reader and then the dissolved O_2 is measured only once at a fixed time after the start of incubation.This protocol can be directly applied for high-throughput measurements.