Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products
Introduction: Contamination of human cell-processed therapeutic products (hCTPs) with tumorigenic/immortalized cellular impurities is a major concern in the manufacturing and quality control of hCTPs. The cellular immortality test based on cell growth analysis is a method for detecting tumorigenic/i...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | Regenerative Therapy |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320422001067 |
| _version_ | 1811188697597476864 |
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| author | Takamasa Hirai Ken Kono Shinji Kusakawa Satoshi Yasuda Rumi Sawada Akihiko Morishita Shinko Hata Atsushi Wakita Takayasu Kageyama Ryo Takahashi Sono Watanabe Norihiko Shiraishi Yoji Sato |
| author_facet | Takamasa Hirai Ken Kono Shinji Kusakawa Satoshi Yasuda Rumi Sawada Akihiko Morishita Shinko Hata Atsushi Wakita Takayasu Kageyama Ryo Takahashi Sono Watanabe Norihiko Shiraishi Yoji Sato |
| author_sort | Takamasa Hirai |
| collection | DOAJ |
| description | Introduction: Contamination of human cell-processed therapeutic products (hCTPs) with tumorigenic/immortalized cellular impurities is a major concern in the manufacturing and quality control of hCTPs. The cellular immortality test based on cell growth analysis is a method for detecting tumorigenic/immortalized cellular impurities in hCTPs. However, the performance of the cellular immortality test has not yet been well characterized. In this study, we examined the reproducibility of the cellular immortality test in detecting HeLa cells as a model of tumorigenic cellular impurities, as well as the applicability of other models of cellular impurities with different tumorigenicity to the cellular immortality test. Methods: Using HeLa cells as a model for cellular impurities, we measured the growth rate of human mesenchymal stem cells (hMSCs) supplemented with HeLa cells at concentrations ranging from 0.01 to 0.0001% at each passage in three laboratories and evaluated the reproducibility of the detection of immortalized cellular impurities. In addition, HEK293 cells (another immortalized cell line) and MRC-5 cells (a non-immortalized cell line) were employed as cellular impurity models that exhibit different growth characteristics from HeLa cells, and the ability of the cellular immortality test to detect these different impurities when mixed with hMSCs was examined. Results: In the multisite study, the growth rate of hMSCs supplemented with 1 and 10 HeLa cells (0.0001% and 0.001%) significantly increased and reached a plateau in all three laboratories, whereas those of hMSCs alone eventually decreased. Moreover, when hMSCs were supplemented with 10 and 100 HEK293 and MRC-5 cells (0.001% and 0.01%), the growth rate significantly increased. The growth rate of hMSCs supplemented with HEK293 cells increased with passage and remained high, whereas that of hMSCs supplemented with MRC-5 cells eventually decreased, as in the case of hMSCs alone. Conclusions: These results indicate that the cellular immortality test is reproducible and can detect immortalized (i.e., potentially tumorigenic) cells such as HEK293 cells with a lower growth rate than HeLa cells by discriminating against normal cells, which could contribute to ensuring the safety and quality of hCTPs. |
| first_indexed | 2024-04-11T14:22:47Z |
| format | Article |
| id | doaj.art-e9d27fb61bcb42359e4bdeae71d42de3 |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-04-11T14:22:47Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-e9d27fb61bcb42359e4bdeae71d42de32022-12-22T04:19:00ZengElsevierRegenerative Therapy2352-32042022-12-0121540546Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic productsTakamasa Hirai0Ken Kono1Shinji Kusakawa2Satoshi Yasuda3Rumi Sawada4Akihiko Morishita5Shinko Hata6Atsushi Wakita7Takayasu Kageyama8Ryo Takahashi9Sono Watanabe10Norihiko Shiraishi11Yoji Sato12Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, Japan; Department of Quality Assurance Science for Pharmaceuticals, Graduate School of Pharmaceutical Sciences, Nagoya City University, Aichi, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, JapanIg-M Co., Hyogo, JapanIg-M Co., Hyogo, JapanClinical Pathology Division, Tsukuba Research Institute, BoZo Research Center Inc., Ibaraki, JapanClinical Pathology Division, Tsukuba Research Institute, BoZo Research Center Inc., Ibaraki, JapanClinical Pathology Division, Tsukuba Research Institute, BoZo Research Center Inc., Ibaraki, JapanAnalytical Research Group, Research Division, HEALIOS K.K., Hyogo, JapanNew Healthcare Solutions, Corporate Strategy Department, Strategy Division, Kyowakirin Co., Ltd., Tokyo, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, Japan; Next Generation Life Science Technology Development Project, Kanagawa Institute of Industrial Science and Technology, Kanagawa, Japan; Department of Cellular and Gene Therapy Products, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Corresponding author. Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki Ward, Kawasaki City, Kanagawa 210-9501, Japan. Fax: +81-44-270-6526.Introduction: Contamination of human cell-processed therapeutic products (hCTPs) with tumorigenic/immortalized cellular impurities is a major concern in the manufacturing and quality control of hCTPs. The cellular immortality test based on cell growth analysis is a method for detecting tumorigenic/immortalized cellular impurities in hCTPs. However, the performance of the cellular immortality test has not yet been well characterized. In this study, we examined the reproducibility of the cellular immortality test in detecting HeLa cells as a model of tumorigenic cellular impurities, as well as the applicability of other models of cellular impurities with different tumorigenicity to the cellular immortality test. Methods: Using HeLa cells as a model for cellular impurities, we measured the growth rate of human mesenchymal stem cells (hMSCs) supplemented with HeLa cells at concentrations ranging from 0.01 to 0.0001% at each passage in three laboratories and evaluated the reproducibility of the detection of immortalized cellular impurities. In addition, HEK293 cells (another immortalized cell line) and MRC-5 cells (a non-immortalized cell line) were employed as cellular impurity models that exhibit different growth characteristics from HeLa cells, and the ability of the cellular immortality test to detect these different impurities when mixed with hMSCs was examined. Results: In the multisite study, the growth rate of hMSCs supplemented with 1 and 10 HeLa cells (0.0001% and 0.001%) significantly increased and reached a plateau in all three laboratories, whereas those of hMSCs alone eventually decreased. Moreover, when hMSCs were supplemented with 10 and 100 HEK293 and MRC-5 cells (0.001% and 0.01%), the growth rate significantly increased. The growth rate of hMSCs supplemented with HEK293 cells increased with passage and remained high, whereas that of hMSCs supplemented with MRC-5 cells eventually decreased, as in the case of hMSCs alone. Conclusions: These results indicate that the cellular immortality test is reproducible and can detect immortalized (i.e., potentially tumorigenic) cells such as HEK293 cells with a lower growth rate than HeLa cells by discriminating against normal cells, which could contribute to ensuring the safety and quality of hCTPs.http://www.sciencedirect.com/science/article/pii/S2352320422001067Cellular immortality testCell growth analysisCellular therapyTumorigenicityQuality and safety |
| spellingShingle | Takamasa Hirai Ken Kono Shinji Kusakawa Satoshi Yasuda Rumi Sawada Akihiko Morishita Shinko Hata Atsushi Wakita Takayasu Kageyama Ryo Takahashi Sono Watanabe Norihiko Shiraishi Yoji Sato Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products Regenerative Therapy Cellular immortality test Cell growth analysis Cellular therapy Tumorigenicity Quality and safety |
| title | Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products |
| title_full | Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products |
| title_fullStr | Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products |
| title_full_unstemmed | Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products |
| title_short | Evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell-processed therapeutic products |
| title_sort | evaluation of the reproducibility and positive controls of cellular immortality test for the detection of immortalized cellular impurities in human cell processed therapeutic products |
| topic | Cellular immortality test Cell growth analysis Cellular therapy Tumorigenicity Quality and safety |
| url | http://www.sciencedirect.com/science/article/pii/S2352320422001067 |
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