bioGenous™ Mouse Intestinal Organoid Kit with Wnt Optimization is a flexible and efficient serum-free culture system designed for the robust establishment and long-term maintenance of mouse intestinal organoids. Compared with the previously released Mouse Intestinal Organoid Kit (Cat# K2001-MI), this upgraded version independently provides an optional Wnt ligand supplement (Component D), enabling researchers to adjust Wnt signaling intensity according to specific experimental requirements. The Wnt Supplement (Component D) can be selectively included during critical proliferative stages such as early stem cell expansion or organoid establishment from single cells to enhance crypt budding, stemness maintenance, and survival efficiency. When Paneth cells are absent or endogenous Wnt secretion is limited, supplementation with Component D effectively supports early organoid formation. Once stable organoid structures have developed, removing Component D promotes differentiation into mature intestinal organoids composed of LGR5+ intestinal stem cells, KI67+ proliferative cells, ALPi+ absorptive enterocytes, ATOH1+ Paneth cells, and MUC2+ goblet cells.