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Mutation accumulation occurs in hematopoietic stem cells (HSCs) after chemotherapy, which may explain the increased risk for developing a second cancer. However, the cause of increased mutagenesis is unclear. In preliminary data, no clear indications of a “chemotherapy signature”, the mutational profile after chemotherapy, can be observed. We aim to study the effects of different chemotherapy treatments over time on healthy HSCs derived from human umbilical cord blood in vitro.