Data from Effects and Eradication of <i>Mycoplasma</i> Contamination on Patient-derived Colorectal Cancer Organoid Cultures

Abstract

<div><p>Patient-derived organoids are a useful platform for identification and testing of novel precision oncology approaches. Patient-derived organoids are generated by direct culture of patient samples. However, prior to development into patient-derived organoids, these samples are often processed for clinical use, opening the potential for contamination by <i>Mycoplasma</i> and other microbes. While most microbes can be detected by visual inspection, <i>Mycoplasma</i> can go undetected and have substantial impacts on assay results. Given the increased use of patient-derived organoids, there is a growing need for a standardized protocol to detect and remove <i>Mycoplasma</i> from organoid models. In the current study, we report a procedure for <i>Mycoplasma</i> removal by passaging organoids through mice as patient-derived organoid xenografts. <i>In vivo</i> passage of patient-derived organoids followed by re-establishment was 100% effective at decontaminating colorectal patient-derived organoids (<i>n</i> = 9), based on testing with the Sigma LookOut Mycoplasma PCR Detection Kit. This process can serve as a method to re-establish contaminated patient-derived organoids, which represent precious models to study patient-specific genomic features and treatment responses.</p>Significance:<p>Organoids are valuable models of cancer. <i>Mycoplasma</i> contamination can alter organoid drug sensitivity, so there is a need for a standardized protocol to detect and remove <i>Mycoplasma</i> from organoids. We report a simple procedure for removing <i>Mycoplasma</i> from organoids via <i>in vivo</i> passaging through mice followed by re-establishment of organoids.</p></div>