The KhES-3 cell line was established at and provided by Kyoto University (Nakatsuji, 2005 (link)). The protocol of this study was reviewed by the Ethics Committee of CiRA in accordance with the "Guidelines for Derivation and Utilization of Human Embryonic Stem Cells" by the Ministry of Education, Culture, Sports, Science and Technology, Japan. The iPS cell lines were established from healthy Japanese donors at CiRA, Kyoto University, and were approved for use by the Ethics Committee of Kyoto University.
Since it has been reported that the toxicity of antioxidants such as catechin is suppressed in the presence of albumin (Zhang et al., 2016 (link)), maintenance culture was carried out for all cell lines including human ES cells using albumin-free Essential 8 Medium (Thermo Fisher Scientific) in six-well feeder-free culture dishes coated with 5 μg/mL vitronectin (VTN-N; Thermo Fisher Scientific). When seeding the cells, 10 μM CultureSure® Y-27632 (FUJIFILM WAKO) was added, and medium exchange on day 1 and thereafter was performed without Y-27632.
Since it has been reported that the toxicity of antioxidants such as catechin is suppressed in the presence of albumin (Zhang et al., 2016 (link)), maintenance culture was carried out for all cell lines including human ES cells using albumin-free Essential 8 Medium (Thermo Fisher Scientific) in six-well feeder-free culture dishes coated with 5 μg/mL vitronectin (VTN-N; Thermo Fisher Scientific). When seeding the cells, 10 μM CultureSure® Y-27632 (FUJIFILM WAKO) was added, and medium exchange on day 1 and thereafter was performed without Y-27632.
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