用于优化高效培养测定的多站点研究，该培养测定用于体外检测细胞治疗产品中混合的残留未分化人类多能干细胞。
Multisite studies for optimization of a highly efficient culture assay used for in vitro detection of residual undifferentiated human pluripotent stem cells intermingled in cell therapy products.
发表日期：2024 Jun
作者：
Takeshi Watanabe, Satoshi Yasuda, Shinji Kusakawa, Takuya Kuroda, Hatsue Furukawa, Mayumi Futamura, Shigekazu Shimizu, Akihiko Morishita, Shinko Hata, Akiko Koeda, Kana Komatsu, Yoji Sato
来源：
Stem Cell Research & Therapy
摘要：
MEASURE2（人类再生医疗产品非临床安全性评估分析方法的多中心评估研究2）是日本的一项实验性公私合作倡议，旨在标准化人类多能干细胞（hPSC）致瘤性评估的测试方法 -衍生细胞治疗产品（CTP）。 MEASURE2 组织了多中心研究，以优化高效培养 (HEC) 测定的方法，这是一种基于培养的敏感体外测定，用于检测 CTP 中残留的未分化 hPSC。在这些多中心研究中，1) 人类诱导多能性集落形成的效率两种不同培养条件下的干细胞 (hiPSC) 以及 2) 使用将 hiPSC 掺入 hiPSC 衍生间充质干细胞的样品评估 hiPSC 富集过程中与各种抗 hPSC 标记物缀合的微珠的分选效率。集落形成效率在 Chroman 1、Emricasan、Polyamines 和 Trans-ISRIB (CEPT) 组合的培养条件下，其显着高于广泛用于 hPSC 存活的 Y-27632。 CEPT 条件下的实验室间差异也比 Y-27632 条件下的小。与抗 Tra-1-60 抗体缀合的微珠的分选效率比其他研究的各种微珠高得多 (>80%)。这些多中心研究的结果预计将有助于提高HEC 测定，以及 hPSC 衍生 CTP 致瘤性风险评估的未来标准化。© 2024 作者。
MEASURE2 (Multisite Evaluation Study on Analytical Methods for Non-clinical Safety Assessment of HUman-derived REgenerative Medical Products 2) is a Japanese experimental public-private partnership initiative that aims to standardize testing methods for tumorigenicity evaluation of human pluripotent stem cell (hPSC)-derived cell therapy products (CTPs). MEASURE2 organized multisite studies to optimize the methodology of the highly efficient culture (HEC) assay, a sensitive culture-based in vitro assay for detecting residual undifferentiated hPSCs in CTPs.In these multisite studies, 1) the efficiency of colony formation by human induced pluripotent stem cells (hiPSCs) under two different culture conditions and 2) the sorting efficiency of microbeads conjugated to various anti-hPSC markers during hiPSC enrichment were evaluated using samples in which hiPSCs were spiked into hiPSC-derived mesenchymal stem cells.The efficiency of colony formation was significantly higher under culture conditions with the combination of Chroman 1, Emricasan, Polyamines, and Trans-ISRIB (CEPT) than with Y-27632, which is widely used for the survival of hPSCs. Between-laboratory variance was also smaller under the condition with CEPT than with Y-27632. The sorting efficiency of microbeads conjugated with the anti-Tra-1-60 antibody was sufficiently higher (>80%) than those of the other various microbeads investigated.Results of these multisite studies are expected to contribute to improvements in the sensitivity and robustness of the HEC assay, as well as to the future standardization of the tumorigenicity risk assessment of hPSC-derived CTPs.© 2024 The Author(s).