Tumorigenicity assay essential for facilitating safety studies of hiPSC-derived cardiomyocytes for clinical application

• Published in: Scientific reports Vol. 9; no. 1; p. 1881
• Main Authors: Ito, Emiko, Miyagawa, Shigeru, Takeda, Maki, Kawamura, Ai, Harada, Akima, Iseoka, Hiroko, Yajima, Shin, Sougawa, Nagako, Mochizuki-Oda, Noriko, Yasuda, Satoshi, Sato, Yoji, Sawa, Yoshiki
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 13.02.2019; Nature Publishing Group
• Subjects: 13; 13/100; 13/31; 13/51; 38; 38/77; 631/532/1360; 631/532/2064/2158; 64; 64/60; Animals; Carcinogenicity Tests - methods; Cardiomyocytes; Cell Differentiation; Cell Proliferation; Cells, Cultured; Congestive heart failure; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Karyotype; Membrane Glycoproteins - metabolism; multidisciplinary; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - transplantation; Neoplasms - etiology; Pluripotency; Rats; Rats, Nude; RNA-Binding Proteins - metabolism; Science; Science (multidisciplinary); Stem cells; Subculture; Transformed cells; Transplantation; Transplantation - adverse effects; Tumorigenicity; Tumors
• ISSN: 2045-2322; 2045-2322
• 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.