Establishment of kidney cancer organoid cultures

This protocol describes an efficient novel method for cultivating kidney cancer organoids, derived directly from human clear cell renal cell carcinomas (ccRCC) or sorted cancer stem cells. The organoids recapitulate tumor-specific characteristics and display inter-tumor heterogeneity. Here we explain how to establish the organoid cultures, maintain them in defined, serum-free medium and use them for patient-specific drug testing. Within 10-14 days, a patient-derived organoid culture is established and can be serially passaged. Compared to other protocols focusing on organoid cultures derived from pediatric kidney malignancies or healthy kidneys, the protocol described here is optimized for adult ccRCC organoid cultures. Organoids established using this protocol can be used to further investigate tumor biology in a complex 3D in vitro system, complement current preclinical model-based drug studies, and advance the development of personalized therapy design