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The L-WRN cell line is a murine fibroblast cell line derived from the L cells, which are mouse fibroblasts originally isolated from connective tissue. L-WRN cells have been engineered to stably express Wnt3a, R-spondin 3, and Noggin. These factors are critical for the growth and maintenance of intestinal organoids and stem cell cultures. The overexpression of these proteins enhances the proliferation and differentiation of intestinal stem cells, making L-WRN cells a valuable tool for studying intestinal biology and disease modeling.
In addition to their application in organoid culture, L-WRN cells serve as a robust model for investigating Wnt signaling pathways. Wnt signaling is pivotal in regulating cell fate, proliferation, and migration during development and in adult tissues. By providing a consistent and controlled source of Wnt3a, R-spondin 3, and Noggin, L-WRN cells facilitate research into the molecular mechanisms underlying these processes. Researchers can use these cells to dissect the roles of these signaling molecules in various biological contexts, including cancer, tissue regeneration, and developmental biology.
Overall, the L-WRN cell line is a powerful tool in biomedical research due to its ability to support the growth of complex three-dimensional cultures and its utility in studying key signaling pathways. Its role in the advancement of intestinal stem cell research and its contributions to our understanding of Wnt signaling highlight its importance in the field of cellular and molecular biology.
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Protein expression: wnt-3A, R-spondin, noggin
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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