論文 - 詳細
| RRC ID | 63069 |
|---|---|
| 著者 | Watanabe T, Yasuda S, Kusakawa S, Kuroda T, Futamura M, Ogawa M, Mochizuki H, Kikkawa E, Furukawa H, Nagaoka M, Sato Y. |
| タイトル | 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. |
| ジャーナル | Cytotherapy |
| Abstract |
BACKGROUND AIMS:The 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. METHODS:MEASURE first validated the detection sensitivity of HEC assay and then investigated the feasibility of magnetic-activated cell sorting (MACS) to improve sensitivity. RESULTS:The 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%. CONCLUSIONS:These 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. |
| 巻・号 | 23(2) |
| ページ | 176-183 |
| 公開日 | 2021-2-1 |
| DOI | 10.1016/j.jcyt.2020.07.009 |
| PII | S1465-3249(20)30796-9 |
| PMID | 32978066 |
| MeSH | Cell Culture Techniques* Cell Differentiation Cell Separation Cell- and Tissue-Based Therapy* Culture Media Humans Induced Pluripotent Stem Cells* Pluripotent Stem Cells* |
| IF | 4.218 |
| リソース情報 | |
| ヒト・動物細胞 | 201B7(HPS0063) |