Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan.
Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kawasaki, Kanagawa, 210-9501, Japan.
Sci Rep. 2019 Feb 13;9(1):1881. doi: 10.1038/s41598-018-38325-5.
Transplantation of cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSC-CMs) is a promising treatment for heart failure, but residual undifferentiated hiPSCs and malignant transformed cells may lead to tumor formation. Here we describe a highly sensitive tumorigenicity assay for the detection of these cells in hiPSC-CMs. The soft agar colony formation assay and cell growth analysis were unable to detect malignantly transformed cells in hiPSC-CMs. There were no karyotypic abnormalities during hiPSCs subculture and differentiation. The hiPSC markers TRA1-60 and LIN28 showed the highest sensitivity for detecting undifferentiated hiPSCs among primary cardiomyocytes. Transplantation of hiPSC-CMs with a LIN28-positive fraction > 0.33% resulted in tumor formation in nude rats, whereas no tumors were formed when the fraction was < 0.1%. These findings suggested that combination of these in vitro and in vivo tumorigenecity assays can verify the safety of hiPSC-CMs for cell transplantation therapy.
源自人诱导多能干细胞（hiPSC-CMs）的心肌细胞（CMs）移植是心力衰竭的一种有前途的治疗方法，但残留的未分化 hiPSC 和恶性转化细胞可能导致肿瘤形成。在这里，我们描述了一种用于检测 hiPSC-CMs 中这些细胞的高灵敏度致瘤性测定法。软琼脂集落形成测定法和细胞生长分析均无法检测 hiPSC-CMs 中的恶性转化细胞。在 hiPSC 传代和分化过程中没有染色体异常。在原代心肌细胞中，hiPSC 标志物 TRA1-60 和 LIN28 对检测未分化 hiPSC 的灵敏度最高。当 LIN28 阳性部分>0.33%的 hiPSC-CMs 移植时，裸鼠会形成肿瘤，而当该部分<0.1%时则不会形成肿瘤。这些发现表明，这些体外和体内致瘤性测定法的组合可验证 hiPSC-CMs 用于细胞移植治疗的安全性。
