Wnt signaling is known to be involved in early development, maintenance and regeneration of stem cells, and in cancer formation. Wnt signaling has also been found to play an important role in the growth and maintenance of these processes. In particular, Wnt3a has been revealed to be an essential niche component for maintaining the proliferation of Lgr5-positive stem cells in intestinal epithelial cells and is used for the production of various digestive organoids such as the small intestine, large intestine, stomach, pancreas and liver. Although Wnt3a has been conventionally used for the culture of gut organoids, it is a fat-soluble protein, so it forms aggregates in serum-free medium and can not exert its activity sufficiently. In 2016, Mihara et al. found that high Wnt3a activity can be maintained by forming a complex with Wnt3a by Afamin, which is one of the components of serum. In addition, by using Afamin and Wnt3a complex for organoid culture, long-term culture of organoid becomes possible. This new medium will result in optimal success for your organoid experiments.
Mihara, et al, eLife (2016) [PMID: 26902720]
Images on the top panels show a bright field of human color organoids. Images on the bottom panels show fluorescent LGR5 positive stem cells that express tdTomato regulated by Lgr5 promoter. Afamin/Wnt3a CM maintained LGR5 positive stem cell growth is seen at greater levels compared with cell growth in purified Wnt3a (300 ng/mL).
Product Code # | Product Name | Main Components | Size | Solvent |
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J2-001 | Afamin/Wnt3a CM | Mouse Wnt3a Human Afamin |
10 mL | Advanced D-MEM/F-12 |
Recombinant Afamin/Wnt3a was purified from the culture supernatant of CHO-K1 cells co-expressing Afamin and Wnt3a.
Product Code # | Product Name | Main Components | Size | Solvent |
---|---|---|---|---|
J2-002 | Recombinant Afamin/Wnt3a | Mouse Wnt3a Human Afamin |
60 µg/300 µL | 20 mM Tris-HCl (pH 7.4), 150 mM NaCl |
An organoid strain expressing tdTomato regulated by Lgr5 promoter was cultured in the presence of Recombinant Afamin/Wnt3a and Recombinant Wnt3a of competitor A. It was confirmed that Recombinant Afamin/Wnt3a maintains LGR5-positive cells even at low concentrations.
This data obtained from collaboration with Dr. Sato, Keio Univ.
Mihara E et al. Active and water-soluble form of lipidated Wnt protein is maintained by a serum glycoprotein afamin/α-albumin, Elife. 2016, 5:e11621 . [PMID: 26902720 ]
→Reference Lists of Afamin/Wnt3a CM