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Lgr5-positive stem cells can be supplemented with the essential growth factors Egf, Noggin, and R-Spondin, which allows us to culture ever-expanding primary 3D epithelial structures  in vitro . Both the architecture and physiological properties of these 'mini-guts', also called organoids, closely resemble their  in vivo  counterparts. This makes them an attractive model system for the small intestinal epithelium. Using retroviral transduction, functional genetics can now be performed by conditional gene overexpression or knockdown. This video demonstrates the procedure of organoid culture, the generation of retroviruses, and the retroviral transduction of organoids to assist phenotypic analysis of the small intestinal epithelium  in vitro . This novel organotypic model system in combination with retroviral mediated gene expression provides a valuable tool for rapid analysis of gene function  in vitro  without the need of costly and time-consuming generation for transgenic animals.

A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture