Details
| Original language | English |
|---|---|
| Article number | 108929 |
| Number of pages | 10 |
| Journal | Biochemical engineering journal |
| Volume | 196 |
| Early online date | 13 Jul 2023 |
| Publication status | Published - Jul 2023 |
| Externally published | Yes |
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 aggregates, 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 functionality. 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.
Keywords
- Adoptive Cell Therapy (ACT), ATMPs (advanced therapy medicinal products), Cytotoxicity, Kinetics, Natural Killer Cells, NK-92
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Environmental Science(all)
- Environmental Engineering
- Engineering(all)
- Biomedical Engineering
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Biochemical engineering journal, Vol. 196, 108929, 07.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Kinetics of NK-92 growth and functionality in pseudo-static cultures
AU - García Aponte, Oscar Fabian
AU - Kozma, Bence
AU - Egger, Dominik
AU - Kasper, Cornelia
AU - Herwig, Christoph
N1 - Funding Information: This work was performed within the framework of the Competence Center CHASE GmbH, funded by the Austrian Research Promotion Agency (grant number 868615 ). The COMET Centre CHASE is funded within the framework of COMET – Competence Centers for Excellent Technologies by BMVIT , BMDW , the Federal Provinces of Upper Austria and Vienna.
PY - 2023/7
Y1 - 2023/7
N2 - 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 aggregates, 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 functionality. 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.
AB - 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 aggregates, 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 functionality. 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.
KW - Adoptive Cell Therapy (ACT)
KW - ATMPs (advanced therapy medicinal products)
KW - Cytotoxicity
KW - Kinetics
KW - Natural Killer Cells
KW - NK-92
UR - http://www.scopus.com/inward/record.url?scp=85152737845&partnerID=8YFLogxK
U2 - 10.1016/j.bej.2023.108929
DO - 10.1016/j.bej.2023.108929
M3 - Article
AN - SCOPUS:85152737845
VL - 196
JO - Biochemical engineering journal
JF - Biochemical engineering journal
SN - 1369-703X
M1 - 108929
ER -