A method for culturing patient-derived lung cancer organoids from surgically resected tissues and biopsy samples

Abstract

Cancer model systems that maintain the genetic and phenotypic characteristics of human cancers are crucial for the study of precision cancer medicine. In this respect, patient-derived cancer organoids have been developed as preclinical models of various human cancers, with significant advantages over previous cancer models including patient-derived xenografts and cell lines. We recently reported a culture system of patient-derived lung cancer organoids (LCOs) that retain the characteristics of patients’ tumors. Here, we describe a detailed protocol for establishing LCOs from surgically resected tumor tissues and endoscopic biopsy samples. This method improves the efficiency of setting up LCOs composed of pure cancer cells and describes an additional procedure for reconstructing LCOs after cryopreservation. We confirmed that stored LCOs had the ability to self-organize and retain the morphological and genetic characteristics of their parental tissues. They also maintained their responsive properties to certain anticancer drugs after thawing. In conclusion, our method efficiently generates LCOs that enable anticancer drug screening at the individual patient level.

Patient-Derived Free-Floating Non-Small Cell Lung Cancer Organoids: A Versatile Tool for Personalized Testing of Chemotherapeutic Drugs

“…49,[52][53][54] Fast assembly of free-floating organoids appears to be important not only for PDTO survival but also for better mimicking tumor microenvironment. It has been revealed that a long-term organoid cultivation often leads to overgrowth of stromal cells like cancer-associated fibroblasts, 61 or eliminating of immune cells due to lack of their expansion. 19 While the deficiency of immune cells can still be compensated for by adding them to tumor organoid culture, 62,63 solving the problem of their contamination with fibroblasts is more difficult.…”