title: Effects of diluents on cell culture viability measured by automated cell counter.
abstract: Commercially available automated cell counters based on trypan blue dye-exclusion are widely used in industrial cell culture process development and manufacturing to increase throughput and eliminate inherent variability in subjective interpretation associated with manual hemocytometers. When using these cell counters, sample dilution is often necessary to stay within the assay measurement range; however, the effect of time and diluents on cell culture is not well understood. This report presents the adverse effect of phosphate buffered saline as a diluent on cell viability when used in combination with an automated cell counter. The reduced cell viability was attributed to shear stress introduced by the automated cell counter. Furthermore, length of time samples were incubated in phosphate buffered saline also contributed to the observed drop in cell viability. Finally, as erroneous viability measurements can severely impact process decisions and product quality, this report identifies several alternative diluents that can maintain cell culture viability over time in order to ensure accurate representation of cell culture conditions.
authors.affiliation: BioProcess Development, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Aaron
authors.initials: A
authors.lastname: Chen
authors.affiliation: BioProcess Development, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Matthew
authors.initials: M
authors.lastname: Leith
authors.affiliation: BioProcess Development, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Roger
authors.initials: R
authors.lastname: Tu
authors.affiliation: BioProcess Development, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Gurpreet
authors.initials: G
authors.lastname: Tahim
authors.affiliation: Clinical Manufacturing, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Anish
authors.initials: A
authors.lastname: Sudra
authors.affiliation: BioProcess Development, Seattle Genetics, Inc., Bothell, Washington, United States of America.
authors.forename: Swapnil
authors.initials: S
authors.lastname: Bhargava
mesh_terms: D000818:Animals; D001331:Automation; D016466:CHO Cells; D002452:Cell Count / Q000379:methods* / Q000592:standards; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003412:Cricetulus; D003470:Culture Media
chemical_list: D003470:Culture Media