Multisite studies for validation and improvement of a highly efficient culture assay for detection of undifferentiated human pluripotent stem cells intermingled in cell therapy products |
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| Authors: | Takeshi Watanabe Satoshi Yasuda Shinji Kusakawa Takuya Kuroda Mayumi Futamura Mitsuhide Ogawa Hidemi Mochizuki Eri Kikkawa Hatsue Furukawa Masato Nagaoka Yoji Sato |
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| Institution: | 1. Drug Safety Research and Evaluation, Takeda Pharmaceutical Company Limited, Fujisawa, Japan;2. The Committee for Non-Clinical Safety Evaluation of Pluripotent Stem Cell-Derived Product, Forum for Innovative Regenerative Medicine, Tokyo, Japan;3. Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kawasaki, Japan;4. Drug Discovery Support Division, Tsukuba Research Institute, BoZo Research Center Inc, Tsukuba, Japan;5. CMIC Bioresearch Center, CMIC Pharma Science Co, Ltd, Hokuto, Japan;6. Research Planning Section, Ina Research Inc, Ina-shi, Japan;7. Research Division, HEALIOS K.K., Kobe, Japan;8. Integrated & Translational Science, Axcelead Drug Discovery Partners, Inc, Fujisawa, Japan;9. Life Science Research Laboratory, Tosoh Corporation, Ayase-shi, Japan |
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| Abstract: | Background aimsThe Multisite Evaluation Study on Analytical Methods for Non-Clinical Safety Assessment of Human-Derived Regenerative Medical Products (MEASURE) is a Japanese experimental public–private partnership initiative, which aims to standardize methodology for tumorigenicity evaluation of human pluripotent stem cell (hPSC)-derived cell therapy products (CTPs). Undifferentiated hPSCs possess tumorigenic potential, and thus residual undifferentiated hPSCs are one of the major hazards for the risk of tumor formation from hPSC-derived CTPs. Among currently available assays, a highly efficient culture (HEC) assay is reported to be one of the most sensitive for the detection of residual undifferentiated hPSCs.MethodsMEASURE first validated the detection sensitivity of HEC assay and then investigated the feasibility of magnetic-activated cell sorting (MACS) to improve sensitivity.ResultsThe multisite experiments confirmed that the lower limit of detection under various conditions to which the human induced pluripotent stem cell lines and culture medium/substrate were subjected was 0.001%. In addition, MACS concentrated cells expressing undifferentiated cell markers and consequently achieved a detection sensitivity of 0.00002%.ConclusionsThese results indicate that HEC assay is highly sensitive and robust and that the application of MACS on this assay is a promising tool for further mitigation of the potential tumorigenicity risk of hPSC-derived CTPs. |
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