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Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing

Authors
 Seul Lee  ;  Yunjoo Joo  ;  Eun Ji Lee  ;  Youngseon Byeon  ;  Jae-Hwan Kim  ;  Kyoung-Ho Pyo  ;  Young Seob Kim  ;  Sun Min Lim  ;  Peter Kilbride  ;  Rohin K Iyer  ;  Mingming Li  ;  Mandy C French  ;  Jung-Yub Lee  ;  Jeeheon Kang  ;  Hyesin Byun  ;  Byoung Chul Cho 
Citation
 PLOS ONE, Vol.19(2) : e0294857, 2024-02 
Journal Title
PLOS ONE
Issue Date
2024-02
MeSH
Cell Line ; Cell Survival ; Cryopreservation* ; Cryoprotective Agents / pharmacology ; Dimethyl Sulfoxide* / pharmacology ; Humans ; Killer Cells, Natural
Abstract
Natural killer (NK) cells have recently shown renewed promise as therapeutic cells for use in treating hematologic cancer indications. Despite this promise, NK cell manufacturing workflows remain largely manual, open, and disconnected, and depend on feeders, as well as outdated unit operations or processes, often utilizing research-grade reagents. Successful scale-up of NK cells critically depends on the availability and performance of nutrient-rich expansion media and cryopreservation conditions that are conducive to high cell viability and recovery post-thaw. In this paper we used Cytiva hardware and media to expand the NK92 cell line in a model process that is suitable for GMP and clinical manufacturing of NK cells. We tested a range of cryopreservation factors including cooling rate, a range of DMSO-containing and DMSO-free cryoprotectants, ice nucleation, and cell density. Higher post-thaw recovery was seen in cryobags over cryovials cooled in identical conditions, and cooling rates of 1°C/min or 2°C/min optimal for cryopreservation in DMSO-containing and DMSO-free cryoprotectants respectively. Higher cell densities of 5x107 cells/ml gave higher post-thaw viability than those cryopreserved at either 1x106 or 5x106 cells/ml. This enabled us to automate, close and connect unit operations within the workflow while demonstrating superior expansion and cryopreservation of NK92 cells. Cellular outputs and performance were conducive to clinical dosing regimens, serving as a proof-of-concept for future clinical and commercial manufacturing. Copyright: © 2024 Lee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Files in This Item:
T202405526.pdf Download
DOI
38394177
Appears in Collections:
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Lim, Sun Min(임선민)
Cho, Byoung Chul(조병철) ORCID logo https://orcid.org/0000-0002-5562-270X
Pyo, Kyoung Ho(표경호) ORCID logo https://orcid.org/0000-0001-5428-0288
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/200568
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