OPTIMIZATION OF PATIENT-DERIVED COLORECTAL CANCER ORGANOIDS PROTOCOLS FOR PERSONALIZED DRUG SCREENINGS.

OBJECTIVES: Recently, the development of technologies to culture intestinal epithelial cells in vitro as different forms of intestinal organoids causes drawing attention in this area. The isolation and culture of colorectal crypts were first demonstrated in the very recent decade with the identification of stem cell marker Lgr5. The growth of crypts into organoids provides an in vitro model for studying the mucosal physiology, colorectal cancer tumorigenesis, and regeneration. In the current study, cancer stem cell isolation, tissue decontamination, organoid freezing and thawing processes, and organoid culture media ingredients have been evaluated. The aim of this investigation is optimization and production of human colorectal cancer organoids. MATERIAL & METHODS: The colorectal cancerous tissues are collected from Dokuz Eylul University Hospital. Isolation method of cancer cell is performed by EDTA solution and agitation is provided by shaker. Matrigel is used to create optimal 3D structure to grow cancer stem cells up to colorectal tissue. L-WRN cell line has been used to produce secreted factors which are WNT-3A, Rspondin and Noggin in purpose of supporting the development of organoids. RESULTS: Different parameters have been evaluated to obtain more efficient organoids. The experiment is performed in our laboratory presented that, human colorectal cancer organoids have been generated and manipulated in our laboratory. CONCLUSIONS: The currency protocol explains how the generation of 3D organoid cultures from cancerous human colorectal tissue is manipulated. The in vitro 3D organoid model is a promising and novel approach which present us more real-like working material. [ABSTRACT FROM AUTHOR]