Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing.
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NK cell manufacturing workflows remain largely manual, open, and disconnected, and depend on feeders, as well as outdated unit operations or pr...
| المؤلفون الرئيسيون: | , , , , , , , , , , , , , , , |
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| التنسيق: | مقال |
| اللغة: | English |
| منشور في: |
Public Library of Science (PLoS)
2024-01-01
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| سلاسل: | PLoS ONE |
| الوصول للمادة أونلاين: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0294857&type=printable |
| _version_ | 1827339644170141696 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Seul Lee |
| collection | DOAJ |
| description | 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NK cell manufacturing workflows remain largely manual, open, and disconnected, and depend on feeders, as well as outdated unit operations or processes, often utilizing research-grade reagents.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. |
| first_indexed | 2024-03-07T20:03:37Z |
| format | Article |
| id | doaj.art-f53e29796dd1462dbf5dea557ec090d1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-03-07T20:03:37Z |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f53e29796dd1462dbf5dea557ec090d12024-02-28T05:31:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-01192e029485710.1371/journal.pone.0294857Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing.Seul LeeYunjoo JooEun Ji LeeYoungseon ByeonJae-Hwan KimKyoung-Ho PyoYoung Seob KimSun Min LimPeter KilbrideRohin K IyerMingming LiMandy C FrenchJung-Yub LeeJeeheon KangHyesin ByunByoung Chul ChoNatural 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0294857&type=printable |
| spellingShingle | 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 Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. PLoS ONE |
| title | Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. |
| title_full | Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. |
| title_fullStr | Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. |
| title_full_unstemmed | Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. |
| title_short | Successful expansion and cryopreservation of human natural killer cell line NK-92 for clinical manufacturing. |
| title_sort | successful expansion and cryopreservation of human natural killer cell line nk 92 for clinical manufacturing |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0294857&type=printable |
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