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Aponte, OFG; Kozma, B; Egger, D; Kasper, C; Herwig, C.
(2023): Kinetics of NK-92 growth and functionality in pseudo-static cultures
BIOCHEM ENG J. 2023; 196, 108929 FullText FullText_BOKU

Abstract:: Understanding the growth and functionality variation of lymphocytes during expansion is fundamental for process control and endpoint determination. However, monitoring CQAs along lymphocyte expansion requires offline cell characterization, hindering timely identification of process deviations and transitions. We developed a culturing protocol that allowed to investigate day-to-day variation of NK-92 cells' cytotoxicity against K-562 cells. Replacing 20% of the culture with fresh medium every day while disrupting the formed lymphocyte ag-gregates, morphological and functionality changes of the NK-92 cells were induced. Our experiments revealed that the adaptation to the pseudo-static protocol divided the culture to two distinctive phases with exponential growth in each. In between, there was an intermittent drop in viability. Significant functionality fluctuations were observed, despite the regular stimulation strategy. Scale-independent descriptors, such as growth rate, were calculated for each phase and the whole culture and correlated with cell morphology parameters and func-tionality. A novel correlation was found between, the cell size-derived, biomass volume and cytotoxicity that was stronger during the first phase, but it was apparent throughout the entire culture. This was possible to capture, only because of the day-to-day monitoring of cytotoxicity. By utilizing the pseudo-static culturing protocol and the methodology, variables, such as biomass volume, were discovered and by this, we are getting closer to find performance indicators suitable to monitor CQAs in a straightforward, non-invasive manner.
Authors BOKU Wien:: Egger Dominik; Kasper Cornelia
Find related publications in this database (Keywords): Natural Killer Cells; NK-92; Cytotoxicity; Kinetics; Adoptive Cell Therapy (ACT); ATMPs (advanced therapy medicinal products)