Refurbishing single use extracorporeal membrane oxygenation (ECMO) oxygenators for in vitro research applications is common. However, the refurbishment protocols that are established in respective laboratories have never been evaluated. In the present study, we aim at proving the relevance of a well-designed refurbishing protocol by quantifying the burden of repeatedly reused oxygenators. We used the same three oxygenators in 5 days of 6 hours whole blood experiments. During each experiment day, the performance of the oxygenators was measured through the evaluation of gas transfer. Between experiment days, each oxygenator was refurbished applying three alternative refurbishment protocols based on purified water, pepsin and citric acid, and hydrogen peroxide solutions, respectively. After the last experiment day, we disassembled the oxygenators for visual inspection of the fiber mats. The refurbishment protocol based on purified water showed strong degeneration with a 40-50 %-performance drop and clearly visible debris on the fiber mats. Hydrogen peroxide performed better; nevertheless, it suffered a 20% decrease in gas transfer as well as clearly visible debris. Pepsin/citric acid performed best in the field, but also suffered from 10% performance loss and very few, but visible debris. The study showed the relevance of a well-suited and well-designed refurbishment protocol. The distinct debris on the fiber mats also suggests that reusing oxygenators is ill-advised for many experiment series, especially regarding hemocompatibility and in vivo testing. Most of all, this study revealed the relevance of stating the status of test oxygenators and, if refurbished, comment on the implemented refurbishment protocol in detail.